Aadc polynucleotides for the treatment of parkinson&#39;s disease

ABSTRACT

The disclosure relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides encoding AADC for the treatment of Parkinson&#39;s Disease.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application is a continuation application which claims thebenefit of U.S. patent application Ser. No. 16/184,466, filed Nov. 8,2018, entitled AADC POLYNUCLEOTIDES FOR THE TREATMENT OF PARKINSON'SDISEASE; which claims priority as continuation application toInternational Application No. PCT/US2018/037437, filed Jun. 14, 2018,entitled AADC POLYNUCLEOTIDES FOR THE TREATMENT OF PARKINSON'S DISEASE;which claims priority to U.S. Provisional Application No. U.S.62/554,155, filed Sep. 5, 2017, entitled AADC POLYNUCLEOTIDES FOR THETREATMENT OF PARKINSON'S DISEASE, and to U.S. Provisional ApplicationNo. U.S. 62/520,084, filed Jun. 15, 2017, entitled AADC POLYNUCLEOTIDESFOR THE TREATMENT OF PARKINSON'S DISEASE; the contents of each beingincorporated by reference herein in their entirety.

REFERENCE TO SEQUENCE LISTING

The present application is being filed along with a Sequence Listing inelectronic format. The Sequence Listing file, entitled 20571042USCON2SL,was created on Jul. 26, 2019, and is 6,421,749 bytes in size. Theinformation in electronic format of the Sequence Listing is incorporatedherein by reference in its entirety.

FIELD OF THE INVENTION

The invention relates to compositions, particularly nucleic acidmolecules, e.g., polynucleotides encoding AADC, for use in the treatmentof Parkinson's disease. In some embodiments such AADC polynucleotidesmay be encoded by or within recombinant adeno-associated viruses (AAVs).

BACKGROUND

Aromatic L-amino acid decarboxylase (AADC) is a homodimeric pyridoxalphosphate-dependent enzyme responsible for the synthesis of dopamine andserotonin. The encoded protein catalyzes the decarboxylation ofL-3,4-dihydroxyphenylalanine (L-DOPA or levodopa) to dopamine;L-5-hydroxytryptophan to serotonin; and L-tryptophan to tryptamine.Defects in this gene are the cause of aromatic L-amino-aciddecarboxylase deficiency (AADCD), which is an inborn error inneurotransmitter metabolism leading to combined serotonin andcatecholamine deficiency that results in severe motor and autonomicdysfunctions.

Parkinson's Disease (PD) is a progressive neurodegenerative disease ofthe central nervous system (CNS) producing sensory and motor symptoms.Dopamine replacement (i.e., levodopa) has been the standardpharmacotherapy for motor impairment in PD. However, the benefit ofdopamine therapy becomes less marked over time, due, in part, to theprogressive death of dopamine-generating cells and corresponding loss ofAADC activity. Furthermore, systemic administration of high-dosedopamine is complicated by side effects, such as fluctuations in motorperformance, dyskinesias, and hallucinations, resulting fromdopaminergic stimulation of the mesolimbic system. One strategy torestore dopaminergic function and minimize side effects is the use ofgene therapy to deliver AADC directly to a targeted region of the CNS.

The adeno-associated virus (AAV) has emerged as an attractive vector forgene therapy due to its long-term gene expression, the inability toautonomously replicate without a helpervirus, the ability to transducedividing and non-diving cells, and the lack of pathogenicity fromwild-type infections (See e.g., Hadaczek et al. Mol. Ther. 18(8),1458-1461, August 2010). AAV is a helper-dependent DNA parvovirus whichbelongs to the genus Dependovirus.

The present disclosure provides such improved nucleic acid constructs,e.g., polynucleotides, for use with AAV-derived vectors comprising dopacarboxylase (“DDC”) gene sequence which encodes a full-length AADCprotein for the purpose of gene therapy in the treatment of Parkinson'sDisease.

The nucleic acid constructs described herein comprise at least a 5′-ITRand a 3′-ITR, each or both of which may be derived from an AAV,positioned about a DDC gene sequence, as well as additional componentsrequired for gene expression and clone selection.

SUMMARY

Described herein are compositions, methods, processes, kits and devicesfor the design, preparation, manufacture and/or formulation of AADCpolynucleotides.

In some embodiments such AADC polynucleotides may be encoded by orcontained within plasmids or vectors or recombinant adeno-associatedviruses (AAV).

The details of various embodiments of the disclosure are set forth inthe description below. Other features, objects, and advantages of thedisclosure will be apparent from the description and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

The foregoing and other objects, features and advantages will beapparent from the following description of particular embodiments of thedisclosure, as illustrated in the accompanying drawings. The drawingsare not necessarily to scale, emphasis instead being placed uponillustrating the principles of various embodiments of the disclosure.

FIG. 1 is a schematic of a viral genome of the disclosure.

DETAILED DESCRIPTION I. Compositions Adeno-Associated Viruses (AAVs) andAAV Particles

Viruses of the Parvoviridae family are small non-enveloped icosahedralcapsid viruses characterized by a single stranded DNA genome.Parvoviridae family viruses consist of two subfamilies: Parvovirinae,which infect vertebrates, and Densovirinae, which infect invertebrates.Due to its relatively simple structure, easily manipulated usingstandard molecular biology techniques, this virus family is useful as abiological tool. The genome of the virus may be modified to contain aminimum of components for the assembly of a functional recombinantvirus, or viral particle, which is loaded with or engineered to expressor deliver a desired payload, which may be delivered to a target cell,tissue, organ, or organism.

The parvoviruses and other members of the Parvoviridae family aregenerally described in Kenneth I. Berns, “Parvoviridae: The Viruses andTheir Replication,” Chapter 69 in FIELDS VIROLOGY (3d Ed. 1996), thecontents of which are incorporated by reference in their entirety.

The Parvoviridae family comprises the Dependovirus genus which includesadeno-associated viruses (AAV) capable of replication in vertebratehosts including, but not limited to, human, primate, bovine, canine,equine, and ovine species.

The vector genome is a linear, single-stranded DNA (ssDNA) moleculeapproximately 5,000 nucleotides (nt) in length. The AAV viral genome cancomprise a payload region and at least one inverted terminal repeat(ITR) or ITR region. ITRs traditionally flank the coding nucleotidesequences for the non-structural proteins (encoded by Rep genes) and thestructural proteins (encoded by capsid genes or Cap genes). While notwishing to be bound by theory, an AAV viral genome typically comprisestwo ITR sequences. The vector genome comprises a characteristic T-shapedhairpin structure defined by the self-complementary terminal 145 nt ofthe 5′ and 3′ ends of the ssDNA which form an energetically stabledouble stranded region. The double stranded hairpin structures comprisemultiple functions including, but not limited to, acting as an originfor DNA replication by functioning as primers for the endogenous DNApolymerase complex of the host viral replication cell.

In addition to the encoded heterologous payload, AAV particles maycomprise the viral genome, in whole or in part, of any naturallyoccurring and/or recombinant AAV serotype nucleotide sequence orvariant. AAV variants may have sequences of significant homology at thenucleic acid (genome or capsid) and amino acid levels (capsids), toproduce constructs which are generally physical and functionalequivalents, replicate by similar mechanisms, and assemble by similarmechanisms. Chiorini et al., J. Vir. 71: 6823-33 (1997); Srivastava etal., J. Vir. 45:555-64 (1983); Chiorini et al., J. Vir. 73:1309-1319(1999); Rutledge et al., J. Vir. 72:309-319 (1998); and Wu et al., J.Vir. 74: 8635-47 (2000), the contents of each of which are incorporatedherein by reference in their entirety.

In one embodiment, AAV particles of the present disclosure arerecombinant AAV particles which are replication defective, lackingsequences encoding functional Rep and Cap proteins within their viralgenome. These defective AAV particles may lack most or all parentalcoding sequences and essentially carry only one or two AAV ITR sequencesand the nucleic acid of interest for delivery to a cell, a tissue, anorgan or an organism.

In one embodiment, the viral genome of the AAV particles of the presentdisclosure comprise at least one control element which provides for thereplication, transcription and translation of a coding sequence encodedtherein. Not all of the control elements need always be present as longas the coding sequence is capable of being replicated, transcribedand/or translated in an appropriate host cell. Non-limiting examples ofexpression control elements include sequences for transcriptioninitiation and/or termination, promoter and/or enhancer sequences,efficient RNA processing signals such as splicing and polyadenylationsignals, sequences that stabilize cytoplasmic mRNA, sequences thatenhance translation efficacy (e.g., Kozak consensus sequence), sequencesthat enhance protein stability, and/or sequences that enhance proteinprocessing and/or secretion.

According to the present disclosure, AAV particles for use intherapeutics and/or diagnostics comprise a virus that has been distilledor reduced to the minimum components necessary for transduction of anucleic acid payload or cargo of interest. In this manner, AAV particlesare engineered as vehicles for specific delivery while lacking thedeleterious replication and/or integration features found in wild-typeviruses.

AAV particles of the present disclosure may be produced recombinantlyand may be based on adeno-associated virus (AAV) parent or referencesequences. As used herein, a “vector” is any molecule or moiety whichtransports, transduces or otherwise acts as a carrier of a heterologousmolecule such as the nucleic acids described herein.

In addition to single stranded AAV particles (e.g., ssAAVs), the presentdisclosure also provides for self-complementary AAV (scAAVs) particles.scAAV particles contain DNA strands which anneal together to form doublestranded DNA. By skipping second strand synthesis, scAAVs allow forrapid expression in the cell.

In one embodiment, the AAV particle of the present disclosure is anscAAV.

In one embodiment, the AAV particle of the present disclosure is anssAAV.

Methods for producing and/or modifying AAV particles are disclosed inthe art such as pseudotyped AAV particles (PCT Patent Publication Nos.WO200028004; WO200123001; WO2004112727; WO 2005005610 and WO 2005072364,the content of each of which is incorporated herein by reference in itsentirety).

AAV particles may be modified to enhance the efficiency of delivery.Such modified AAV particles can be packaged efficiently and be used tosuccessfully infect the target cells at high frequency and with minimaltoxicity. In some embodiments the capsids of the AAV particles areengineered according to the methods described in US Publication NumberUS 20130195801, the contents of which are incorporated herein byreference in their entirety.

In one embodiment, the AAV particles comprising a payload regionencoding the polypeptides of the disclosure may be introduced intomammalian cells.

AAV Serotypes

AAV particles of the present disclosure may comprise or be derived fromany natural or recombinant AAV serotype. According to the presentdisclosure, the AAV particles may utilize or be based on a serotypeselected from any of the following PHP.B, PHP.A, AAV1, AAV2, AAV2G9,AAV3, AAV3a, AAV3b, AAV3-3, AAV4, AAV4-4, AAV5, AAV6, AAV6.1,AAV6.2,AAV6.1.2, AAV7, AAV7.2, AAV8, AAV9, AAV9.11, AAV9.13, AAV9.16, AAV9.24,AAV9.45, AAV9.47, AAV9.61, AAV9.68, AAV9.84, AAV9.9, AAV10, AAV11,AAV12, AAV16.3, AAV24.1, AAV27.3, AAV42.12, AAV42-1b, AAV42-2, AAV42-3a,AAV42-3b, AAV42-4, AAV42-5a, AAV42-5b, AAV42-6b, AAV42-8, AAV42-10,AAV42-11, AAV42-12, AAV42-13, AAV42-15, AAV42-aa, AAV43-1, AAV43-12,AAV43-20, AAV43-21, AAV43-23, AAV43-25, AAV43-5, AAV44.1, AAV44.2,AAV44.5, AAV223.1, AAV223.2, AAV223.4, AAV223.5, AAV223.6, AAV223.7,AAV1-7/rh.48, AAV1-8/rh.49, AAV2-15/rh.62, AAV2-3/rh.61, AAV2-4/rh.50,AAV2-5/rh.51, AAV3.1/hu.6, AAV3.1/hu.9, AAV3-9/rh.52,AAV3-11/rh.53,AAV4-8/r11.64, AAV4-9/rh.54, AAV4-19/rh.55, AAV5-3/rh.57,AAV5-22/rh.58, AAV7.3/hu.7, AAV16.8/hu.10, AAV16.12/hu.11, AAV29.3/bb.1,AAV29.5/bb.2, AAV106.1/hu.37, AAV114.3/hu.40, AAV127.2/hu.41,AAV127.5/hu.42, AAV128.3/hu.44, AAV130.4/hu.48, AAV145.1/hu.53,AAV145.5/hu.54, AAV145.6/hu.55, AAV161.10/hu.60, AAV161.6/hu.61,AAV33.12/hu.17, AAV33.4/hu.15, AAV33.8/hu.16, AAV52/hu.19,AAV52.1/hu.20, AAV58.2/hu.25, AAVA3.3, AAVA3.4, AAVA3.5, AAVA3.7, AAVC1,AAVC2, AAVC5, AAV-DJ, AAV-DJ8, AAVF3, AAVF5, AAVH2, AAVrh.72, AAVhu.8,AAVrh.68, AAVrh.70, AAVpi.1, AAVpi.3, AAVpi.2, AAVrh.60, AAVrh.44,AAVrh.65, AAVrh.55, AAVrh.47, AAVrh.69, AAVrh.45, AAVrh.59, AAVhu.12,AAVH6, AAVLK03, AAVH-1/hu.1, AAVH-5/hu.3, AAVLG-10/rh.40, AAVLG-4/rh.38,AAVLG-9/hu.39, AAVN721-8/rh.43, AAVCh.5, AAVCh.5R1, AAVcy.2, AAVcy.3,AAVcy.4, AAVcy.5, AAVCy.5R1, AAVCy.5R2, AAVCy.5R3, AAVCy.5R4, AAVcy.6,AAVhu.1, AAVhu.2, AAVhu.3, AAVhu.4, AAVhu.5, AAVhu.6, AAVhu.7, AAVhu.9,AAVhu.10, AAVhu.11, AAVhu.13, AAVhu.15, AAVhu.16, AAVhu.17, AAVhu.18,AAVhu.20, AAVhu.21, AAVhu.22, AAVhu.23.2, AAVhu.24, AAVhu.25, AAVhu.27,AAVhu.28, AAVhu.29, AAVhu.29R, AAVhu.31, AAVhu.32, AAVhu.34, AAVhu.35,AAVhu.37, AAVhu.39, AAVhu.40, AAVhu.41, AAVhu.42, AAVhu.43, AAVhu.44,AAVhu.44R1, AAVhu.44R2, AAVhu.44R3, AAVhu.45, AAVhu.46, AAVhu.47,AAVhu.48, AAVhu.48R1, AAVhu.48R2, AAVhu.48R3, AAVhu.49, AAVhu.51,AAVhu.52, AAVhu.54, AAVhu.55, AAVhu.56, AAVhu.57, AAVhu.58, AAVhu.60,AAVhu.61, AAVhu.63, AAVhu.64, AAVhu.66, AAVhu.67, AAVhu.14/9, AAVhu.t19, AAVrh.2, AAVrh.2R, AAVrh.8, AAVrh.8R, AAVrh.10, AAVrh.12, AAVrh.13,AAVrh.13R, AAVrh.14, AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.20, AAVrh.21,AAVrh.22, AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.31, AAVrh.32, AAVrh.33,AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37, AAVrh.37R2, AAVrh.38, AAVrh.39,AAVrh.40, AAVrh.46, AAVrh.48, AAVrh.48.1, AAVrh.48.1.2, AAVrh.48.2,AAVrh.49, AAVrh.51, AAVrh.52, AAVrh.53, AAVrh.54, AAVrh.56, AAVrh.57,AAVrh.58, AAVrh.61, AAVrh.64, AAVrh.64R1, AAVrh.64R2, AAVrh.67,AAVrh.73, AAVrh.74, AAVrh8R, AAVrh8R A586R mutant, AAVrh8R R533A mutant,AAAV, BAAV, caprine AAV, bovine AAV, ovine AAV, AAVhE1.1, AAVhEr1.5,AAVhER1.14, AAVhEr1.8, AAVhEr1.16, AAVhEr1.18, AAVhEr1.35, AAVhEr1.7,AAVhEr1.36, AAVhEr2.29, AAVhEr2.4, AAVhEr2.16, AAVhEr2.30, AAVhEr2.31,AAVhEr2.36, AAVhER1.23, AAVhEr3.1, AAV2.5T, AAV-PAEC, AAV-LK01,AAV-LK02, AAV-LK03, AAV-LK04, AAV-LK05, AAV-LK06, AAV-LK07, AAV-LK08,AAV-LK09, AAV-LK10, AAV-LK11, AAV-LK12, AAV-LK13, AAV-LK14, AAV-LK15,AAV-LK16, AAV-LK17, AAV-LK18, AAV-LK19, AAV-PAEC2, AAV-PAEC4, AAV-PAEC6,AAV-PAEC7, AAV-PAEC8, AAV-PAEC11, AAV-PAEC12, AAV-2-pre-miRNA-101,AAV-8h, AAV-8b, AAV-h, AAV-b, AAV SM 10-2, AAV Shuffle 100-1, AAVShuffle 100-3, AAV Shuffle 100-7, AAV Shuffle 10-2, AAV Shuffle 10-6,AAV Shuffle 10-8, AAV Shuffle 100-2, AAV SM 10-1, AAV SM 10-8, AAV SM100-3, AAV SM 100-10, BNP61 AAV, BNP62 AAV, BNP63 AAV, AAVrh.50,AAVrh.43, AAVrh.62, AAVrh.48, AAVhu.19, AAVhu.11, AAVhu.53,AAV4-8/rh.64, AAVLG-9/hu.39, AAV54.5/hu.23, AAV54.2/hu.22,AAV54.7/hu.24, AAV54.1/hu.21, AAV54.4R/hu.27, AAV46.2/hu.28,AAV46.6/hu.29, AAV128.1/hu.43, true type AAV (ttAAV), UPENN AAV 10,Japanese AAV 10 serotypes, AAV CBr-7.1, AAV CBr-7.10, AAV CBr-7.2, AAVCBr-7.3, AAV CBr-7.4, AAV CBr-7.5, AAV CBr-7.7, AAV CBr-7.8, AAVCBr-B7.3, AAV CBr-B7.4, AAV CBr-E1, AAV CBr-E2, AAV CBr-E3, AAV CBr-E4,AAV CBr-E5, AAV CBr-e5, AAV CBr-E6, AAV CBr-E7, AAV CBr-E8, AAV CHt-1,AAV CHt-2, AAV CHt-3, AAV CHt-6.1, AAV CHt-6.10, AAV CHt-6.5, AAVCHt-6.6, AAV CHt-6.7, AAV CHt-6.8, AAV CHt-P1, AAV CHt-P2, AAV CHt-P5,AAV CHt-P6, AAV CHt-P8, AAV CHt-P9, AAV CKd-1, AAV CKd-10, AAV CKd-2,AAV CKd-3, AAV CKd-4, AAV CKd-6, AAV CKd-7, AAV CKd-8, AAV CKd-B1, AAVCKd-B2, AAV CKd-B3, AAV CKd-B4, AAV CKd-B5, AAV CKd-B6, AAV CKd-B7, AAVCKd-B8, AAV CKd-H1, AAV CKd-H2, AAV CKd-H3, AAV CKd-H4, AAV CKd-H5, AAVCKd-H6, AAV CKd-N3, AAV CKd-N4, AAV CKd-N9, AAV CLg-F1, AAV CLg-F2, AAVCLg-F3, AAV CLg-F4, AAV CLg-F5, AAV CLg-F6, AAV CLg-F7, AAV CLg-F8, AAVCLv-1, AAV CLv1-1, AAV Clv1-10, AAV CLv1-2, AAV CLv-12, AAV CLv1-3, AAVCLv-13, AAV CLv1-4, AAV Clv1-7, AAV Clv1-8, AAV Clv1-9, AAV CLv-2, AAVCLv-3, AAV CLv-4, AAV CLv-6, AAV CLv-8, AAV CLv-D1, AAV CLv-D2, AAVCLv-D3, AAV CLv-D4, AAV CLv-D5, AAV CLv-D6, AAV CLv-D7, AAV CLv-D8, AAVCLv-E1, AAV CLv-K1, AAV CLv-K3, AAV CLv-K6, AAV CLv-L4, AAV CLv-L5, AAVCLv-L6, AAV CLv-M1, AAV CLv-M11, AAV CLv-M2, AAV CLv-M5, AAV CLv-M6, AAVCLv-M7, AAV CLv-M8, AAV CLv-M9, AAV CLv-R1, AAV CLv-R2, AAV CLv-R3, AAVCLv-R4, AAV CLv-R5, AAV CLv-R6, AAV CLv-R7, AAV CLv-R8, AAV CLv-R9, AAVCSp-1, AAV CSp-10, AAV CSp-11, AAV CSp-2, AAV CSp-3, AAV CSp-4, AAVCSp-6, AAV CSp-7, AAV CSp-8, AAV CSp-8.10, AAV CSp-8.2, AAV CSp-8.4, AAVCSp-8.5, AAV CSp-8.6, AAV CSp-8.7, AAV CSp-8.8, AAV CSp-8.9, AAV CSp-9,AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5, AAVF1/HSC1, AAVF11/HSC11,AAVF12/HSC12, AAVF13/HSC13, AAVF14/HSC14, AAVF15/HSC15, AAVF16/HSC16,AAVF17/HSC17, AAVF2/HSC2, AAVF3/HSC3, AAVF4/HSC4, AAVF5/HSC5,AAVF6/HSC6, AAVF7/HSC7, AAVF8/HSC8, AAVF9/HSC9, PHP.B (AAV-PHP.B), PHP.A(AAV.PHP.A), G2B-26, G2B-13, TH1.1-32, TH1.1-35, AAVPHP.B2, AAVPHP.B3,AAVPHP.N/PHP.B-DGT, AAVPHP.B-EST, AAVPHP.B-GGT, AAVPHP.B-ATP,AAVPHP.B-ATT-T, AAVPHP.B-DGT-T, AAVPHP.B-GGT-T, AAVPHP.B-SGS,AAVPHP.B-AQP, AAVPHP.B-QQP, AAVPHP.B-SNP(3), AAVPHP.B-SNP, AAVPHP.B-QGT,AAVPHP.B-NQT, AAVPHP.B-EGS, AAVPHP.B-SGN, AAVPHP.B-EGT, AAVPHP.B-DST,AAVPHP.B-DST, AAVPHP.B-STP, AAVPHP.B-PQP, AAVPHP.B-SQP, AAVPHP.B-QLP,AAVPHP.B-TMP, AAVPHP.B-TTP, AAVPHP.S/G2A12, AAVG2A15/G2A3, AAVG2B4,and/or AAVG2B5, and variants thereof.

In some embodiments, the AAV serotype may be, or have, a modification asdescribed in United States Publication No. US 20160361439, the contentsof which are herein incorporated by reference in their entirety, such asbut not limited to, Y252F, Y272F, Y444F, Y500F, Y700F, Y704F, Y730F,Y275F, Y281F, Y508F, Y576F, Y612G, Y673F, and Y720F of the wild-typeAAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10,AAV11,AAV12, and hybrids thereof.

In some embodiments, the AAV serotype may be, or have, a mutation asdescribed in U.S. Pat. No. 9,546,112, the contents of which are hereinincorporated by reference in their entirety, such as, but not limitedto, at least two, but not all the F129L, D418E, K531E,L584F, V598A andH642N mutations in the sequence of AAV6 (SEQ ID NO:4 of U.S. Pat. No.9,546,112), AAV1 (SEQ ID NO:6 of U.S. Pat. No. 9,546,112), AAV2, AAV3,AAV4, AAV5, AAV7, AAV9, AAV10 or AAV11 or derivatives thereof. In yetanother embodiment, the AAV serotype may be, or have, an AAV6 sequencecomprising the K531E mutation (SEQ ID NO:5 of U.S. Pat. No. 9,546,112).

In some embodiments, the AAV serotype may be, or have, a mutation in theAAV1 sequence, as described in in United States Publication No. US20130224836, the contents of which are herein incorporated by referencein their entirety, such as, but not limited to, at least one of thesurface-exposed tyrosine residues, preferably, at positions 252, 273,445, 701, 705 and 731 of AAV1 (SEQ ID NO: 2 of US 20130224836)substituted with another amino acid, preferably with a phenylalanineresidue. In one embodiment, the AAV serotype may be, or have, a mutationin the AAV9 sequence, such as, but not limited to, at least one of thesurface-exposed tyrosine residues, preferably, at positions 252, 272,444, 500, 700, 704 and 730 of AAV2 (SEQ ID NO: 4 of US 20130224836)substituted with another amino acid, preferably with a phenylalanineresidue. In one embodiment, the tyrosine residue at position 446 of AAV9(SEQ ID NO: 6 US 20130224836) is substituted with a phenylalanineresidue.

In some embodiments, the serotype may be AAV2 or a variant thereof, asdescribed in International Publication No. WO2016130589, hereinincorporated by reference in its entirety. The amino acid sequence ofAAV2 may comprise N587A, E548A, or N708A mutations. In one embodiment,the amino acid sequence of any AAV may comprise a V708K mutation.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in United States Publication No. US20030138772, the contentsof which are herein incorporated by reference in their entirety, suchas, but not limited to, AAV1 (SEQ ID NO: 6 and 64 of US20030138772),AAV2 (SEQ ID NO: 7 and 70 of US20030138772), AAV3 (SEQ ID NO: 8 and 71of US20030138772), AAV4 (SEQ ID NO: 63 of US20030138772), AAV5 (SEQ IDNO: 114 of US20030138772), AAV6 (SEQ ID NO: 65 of US20030138772), AAV7(SEQ ID NO: 1-3 of US20030138772), AAV8 (SEQ ID NO: 4 and 95 ofUS20030138772), AAV9 (SEQ ID NO: 5 and 100 of US20030138772), AAV10 (SEQID NO: 117 of US20030138772), AAV11 (SEQ ID NO:118 of US20030138772),AAV12 (SEQ ID NO: 119 of US20030138772), AAVrh10 (amino acids 1 to 738of SEQ ID NO: 81 of US20030138772), AAV16.3 (US20030138772 SEQ ID NO:10), AAV29.3/bb.1 (US20030138772 SEQ ID NO: 11), AAV29.4 (US20030138772SEQ ID NO: 12), AAV29.5/bb.2 (US20030138772 SEQ ID NO: 13), AAV1.3(US20030138772 SEQ ID NO: 14), AAV13.3 (US20030138772 SEQ ID NO: 15),AAV24.1 (US20030138772 SEQ ID NO: 16), AAV27.3 (US20030138772 SEQ ID NO:17), AAV7.2 (US20030138772 SEQ ID NO: 18), AAVC1 (US20030138772 SEQ IDNO: 19), AAVC3 (US20030138772 SEQ ID NO: 20), AAVC5 (US20030138772 SEQID NO: 21), AAVF1 (US20030138772 SEQ ID NO: 22), AAVF3 (US20030138772SEQ ID NO: 23), AAVF5 (US20030138772 SEQ ID NO: 24), AAVH6(US20030138772 SEQ ID NO: 25), AAVH2 (US20030138772 SEQ ID NO: 26),AAV42-8 (US20030138772 SEQ ID NO: 27), AAV42-15 (US20030138772 SEQ IDNO: 28), AAV42-5b (US20030138772 SEQ ID NO: 29), AAV42-1b (US20030138772SEQ ID NO: 30), AAV42-13 (US20030138772 SEQ ID NO: 31), AAV42-3a(US20030138772 SEQ ID NO: 32), AAV42-4 (US20030138772 SEQ ID NO: 33),AAV42-5a (US20030138772 SEQ ID NO: 34), AAV42-10 (US20030138772 SEQ IDNO: 35), AAV42-3b (US20030138772 SEQ ID NO: 36), AAV42-11 (US20030138772SEQ ID NO: 37), AAV42-6b (US20030138772 SEQ ID NO: 38), AAV43-1(US20030138772 SEQ ID NO: 39), AAV43-5 (US20030138772 SEQ ID NO: 40),AAV43-12 (US20030138772 SEQ ID NO: 41), AAV43-20 (US20030138772 SEQ IDNO: 42), AAV43-21 (US20030138772 SEQ ID NO: 43), AAV43-23 (US20030138772SEQ ID NO: 44), AAV43-25 (US20030138772 SEQ ID NO: 45), AAV44.1(US20030138772 SEQ ID NO: 46), AAV44.5 (US20030138772 SEQ ID NO: 47),AAV223.1 (US20030138772 SEQ ID NO: 48), AAV223.2 (US20030138772 SEQ IDNO: 49), AAV223.4 (US20030138772 SEQ ID NO: 50), AAV223.5 (US20030138772SEQ ID NO: 51), AAV223.6 (US20030138772 SEQ ID NO: 52), AAV223.7(US20030138772 SEQ ID NO: 53), AAVA3.4 (US20030138772 SEQ ID NO: 54),AAVA3.5 (US20030138772 SEQ ID NO: 55), AAVA3.7 (US20030138772 SEQ ID NO:56), AAVA3.3 (US20030138772 SEQ ID NO: 57), AAV42.12 (US20030138772 SEQID NO: 58), AAV44.2 (US20030138772 SEQ ID NO: 59), AAV42-2(US20030138772 SEQ ID NO: 9), or variants thereof.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in United States Publication No. US20150159173, the contentsof which are herein incorporated by reference in their entirety, suchas, but not limited to, AAV2 (SEQ ID NO: 7 and 23 of US20150159173),rh20 (SEQ ID NO: 1 of US20150159173), rh32/33 (SEQ ID NO: 2 ofUS20150159173), rh39 (SEQ ID NO: 3, 20 and 36 of US20150159173), rh46(SEQ ID NO: 4 and 22 of US20150159173), rh73 (SEQ ID NO: 5 ofUS20150159173), rh74 (SEQ ID NO: 6 of US20150159173), AAV6.1 (SEQ ID NO:29 of US20150159173), rh.8 (SEQ ID NO: 41 of US20150159173), rh.48.1(SEQ ID NO: 44 of US20150159173), hu.44 (SEQ ID NO: 45 ofUS20150159173), hu.29 (SEQ ID NO: 42 of US20150159173), hu.48 (SEQ IDNO: 38 of US20150159173), rh54 (SEQ ID NO: 49 of US20150159173), AAV2(SEQ ID NO: 7 of US20150159173), cy.5 (SEQ ID NO: 8 and 24 ofUS20150159173), rh.10 (SEQ ID NO: 9 and 25 of US20150159173), rh.13 (SEQID NO: 10 and 26 of US20150159173), AAV1 (SEQ ID NO: 11 and 27 ofUS20150159173), AAV3 (SEQ ID NO: 12 and 28 of US20150159173), AAV6 (SEQID NO: 13 and 29 of US20150159173), AAV7 (SEQ ID NO: 14 and 30 ofUS20150159173), AAV8 (SEQ ID NO: 15 and 31 of US20150159173), hu.13 (SEQID NO: 16 and 32 of US20150159173), hu.26 (SEQ ID NO: 17 and 33 ofUS20150159173), hu.37 (SEQ ID NO: 18 and 34 of US20150159173), hu.53(SEQ ID NO: 19 and 35 of US20150159173), rh.43 (SEQ ID NO: 21 and 37 ofUS20150159173), rh2 (SEQ ID NO: 39 of US20150159173), rh.37 (SEQ ID NO:40 of US20150159173), rh.64 (SEQ ID NO: 43 of US20150159173), rh.48 (SEQID NO: 44 of US20150159173), ch.5 (SEQ ID NO 46 of US20150159173), rh.67(SEQ ID NO: 47 of US20150159173), rh.58 (SEQ ID NO: 48 ofUS20150159173), or variants thereof including, but not limited to Cy5R1,Cy5R2, Cy5R3, Cy5R4, rh.13R, rh.37R2, rh.2R, rh.8R, rh.48.1, rh.48.2,rh.48.1.2, hu.44R1, hu.44R2, hu.44R3, hu.29R, ch.5R1, rh64R1, rh64R2,AAV6.2, AAV6.1, AAV6.12, hu.48R1, hu.48R2, and hu.48R3.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in U.S. Pat. No. 7,198,951, the contents of which are hereinincorporated by reference in their entirety, such as, but not limitedto, AAV9 (SEQ ID NO: 1-3 of U.S. Pat. No. 7,198,951), AAV2 (SEQ ID NO: 4of U.S. Pat. No. 7,198,951), AAV1 (SEQ ID NO: 5 of U.S. Pat. No.7,198,951), AAV3 (SEQ ID NO: 6 of U.S. Pat. No. 7,198,951), and AAV8(SEQ ID NO: 7 of U.S. Pat. No. 7,198,951).

In some embodiments, the AAV serotype may be, or have, a mutation in theAAV9 sequence as described by N Pulicherla et al. (Molecular Therapy19(6):1070-1078 (2011), herein incorporated by reference in itsentirety), such as but not limited to, AAV9.9, AAV9.11,AAV9.13, AAV9.16,AAV9.24, AAV9.45, AAV9.47, AAV9.61, AAV9.68, AAV9.84.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in U.S. Pat. No. 6,156,303, the contents of which are hereinincorporated by reference in their entirety, such as, but not limitedto, AAV3B (SEQ ID NO: 1 and 10 of U.S. Pat. No. 6,156,303), AAV6 (SEQ IDNO: 2, 7 and 11 of U.S. Pat. No. 6,156,303), AAV2 (SEQ ID NO: 3 and 8 ofU.S. Pat. No. 6,156,303), AAV3A (SEQ ID NO: 4 and 9, of U.S. Pat. No.6,156,303), or derivatives thereof.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in United States Publication No. US20140359799, the contentsof which are herein incorporated by reference in their entirety, suchas, but not limited to, AAV8 (SEQ ID NO: 1 of US20140359799), AAVDJ (SEQID NO: 2 and 3 of US20140359799), or variants thereof.

In some embodiments, the serotype may be AAVDJ or a variant thereof,such as AAVDJ8 (or AAV-DJ8), as described by Grimm et al. (Journal ofVirology 82(12): 5887-5911 (2008), herein incorporated by reference inits entirety). The amino acid sequence of AAVDJ8 may comprise two ormore mutations in order to remove the heparin binding domain (HBD). As anon-limiting example, the AAV-DJ sequence described as SEQ ID NO: 1 inU.S. Pat. No. 7,588,772, the contents of which are herein incorporatedby reference in their entirety, may comprise two mutations: (1) R587Qwhere arginine (R; Arg) at amino acid 587 is changed to glutamine (Q;Gln) and (2) R590T where arginine (R; Arg) at amino acid 590 is changedto threonine (T; Thr). As another non-limiting example, may comprisethree mutations: (1) K406R where lysine (K; Lys) at amino acid 406 ischanged to arginine (R; Arg), (2) R587Q where arginine (R; Arg) at aminoacid 587 is changed to glutamine (Q; Gln) and (3) R590T where arginine(R; Arg) at amino acid 590 is changed to threonine (T; Thr).

In some embodiments, the AAV serotype may be, or have, a sequence ofAAV4 as described in International Publication No. WO1998011244, thecontents of which are herein incorporated by reference in theirentirety, such as, but not limited to AAV4 (SEQ ID NO: 1-20 ofWO1998011244).

In some embodiments, the AAV serotype may be, or have, a mutation in theAAV2 sequence to generate AAV2G9 as described in InternationalPublication No. WO2014144229 and herein incorporated by reference in itsentirety.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in International Publication No. WO2005033321, the contents ofwhich are herein incorporated by reference in their entirety, such as,but not limited to AAV3-3 (SEQ ID NO: 217 of WO2005033321), AAV1 (SEQ IDNO: 219 and 202 of WO2005033321), AAV106.1/hu.37 (SEQ ID No: 10 ofWO2005033321), AAV114.3/hu.40 (SEQ ID No: 11 of WO2005033321),AAV127.2/hu.41 (SEQ ID NO:6 and 8 of WO2005033321), AAV128.3/hu.44 (SEQID No: 81 of WO2005033321), AAV130.4/hu.48 (SEQ ID NO: 78 ofWO2005033321), AAV145.1/hu.53 (SEQ ID No: 176 and 177 of WO2005033321),AAV145.6/hu.56 (SEQ ID NO: 168 and 192 of WO2005033321), AAV16.12/hu.11(SEQ ID NO:: 153 and 57 of WO2005033321), AAV16.8/hu.10 (SEQ ID NO: 156and 56 of WO2005033321), AAV161.10/hu.60 (SEQ ID No: 170 ofWO2005033321), AAV161.6/hu.61 (SEQ ID No: 174 of WO2005033321),AAV1-7/rh.48 (SEQ ID NO: 32 of WO2005033321), AAV1-8/rh.49 (SEQ ID NOs:103 and 25 of WO2005033321), AAV2 (SEQ ID NO: 211 and 221 ofWO2005033321), AAV2-15/rh.62 (SEQ ID No: 33 and 114 of WO2005033321),AAV2-3/rh.61 (SEQ ID NO: 21 of WO2005033321), AAV2-4/rh.50 (SEQ ID No:23 and 108 of WO2005033321), AAV2-5/rh.51 (SEQ ID NO: 104 and 22 ofWO2005033321), AAV3.1/hu.6 (SEQ ID NO: 5 and 84 of WO2005033321),AAV3.1/hu.9 (SEQ ID NO: 155 and 58 of WO2005033321), AAV3-11/rh.53 (SEQID NO: 186 and 176 of WO2005033321), AAV3-3 (SEQ ID NO: 200 ofWO2005033321), AAV33.12/hu.17 (SEQ ID NO:4 of WO2005033321),AAV33.4/hu.15 (SEQ ID No: 50 of WO2005033321), AAV33.8/hu.16 (SEQ ID No:51 of WO2005033321), AAV3-9/rh.52 (SEQ ID NO: 96 and 18 ofWO2005033321), AAV4-19/rh.55 (SEQ ID NO: 117 of WO2005033321), AAV4-4(SEQ ID NO: 201 and 218 of WO2005033321), AAV4-9/rh.54 (SEQ ID NO: 116of WO2005033321), AAV5 (SEQ ID NO: 199 and 216 of WO2005033321),AAV52.1/hu.20 (SEQ ID NO: 63 of WO2005033321), AAV52/hu.19 (SEQ ID NO:133 of WO2005033321), AAV5-22/rh.58 (SEQ ID No: 27 of WO2005033321),AAV5-3/rh.57 (SEQ ID NO: 105 of WO2005033321), AAV5-3/rh.57 (SEQ ID No:26 of WO2005033321), AAV58.2/hu.25 (SEQ ID No: 49 of WO2005033321), AAV6(SEQ ID NO: 203 and 220 of WO2005033321), AAV7 (SEQ ID NO: 222 and 213of WO2005033321), AAV7.3/hu.7 (SEQ ID No: 55 of WO2005033321), AAV8 (SEQID NO: 223 and 214 of WO2005033321), AAVH-1/hu.1 (SEQ ID No: 46 ofWO2005033321), AAVH-5/hu.3 (SEQ ID No: 44 of WO2005033321), AAVhu.1 (SEQID NO: 144 of WO2005033321), AAVhu.10 (SEQ ID NO: 156 of WO2005033321),AAVhu.11 (SEQ ID NO: 153 of WO2005033321), AAVhu.12 (WO2005033321 SEQ IDNO: 59), AAVhu.13 (SEQ ID NO: 129 of WO2005033321), AAVhu.14/AAV9 (SEQID NO: 123 and 3 of WO2005033321), AAVhu.15 (SEQ ID NO: 147 ofWO2005033321), AAVhu.16 (SEQ ID NO: 148 of WO2005033321), AAVhu.17 (SEQID NO: 83 of WO2005033321), AAVhu.18 (SEQ ID NO: 149 of WO2005033321),AAVhu.19 (SEQ ID NO: 133 of WO2005033321), AAVhu.2 (SEQ ID NO: 143 ofWO2005033321), AAVhu.20 (SEQ ID NO: 134 of WO2005033321), AAVhu.21 (SEQID NO: 135 of WO2005033321), AAVhu.22 (SEQ ID NO: 138 of WO2005033321),AAVhu.23.2 (SEQ ID NO: 137 of WO2005033321), AAVhu.24 (SEQ ID NO: 136 ofWO2005033321), AAVhu.25 (SEQ ID NO: 146 of WO2005033321), AAVhu.27 (SEQID NO: 140 of WO2005033321), AAVhu.29 (SEQ ID NO: 132 of WO2005033321),AAVhu.3 (SEQ ID NO: 145 of WO2005033321), AAVhu.31 (SEQ ID NO: 121 ofWO2005033321), AAVhu.32 (SEQ ID NO: 122 of WO2005033321), AAVhu.34 (SEQID NO: 125 of WO2005033321), AAVhu.35 (SEQ ID NO: 164 of WO2005033321),AAVhu.37 (SEQ ID NO: 88 of WO2005033321), AAVhu.39 (SEQ ID NO: 102 ofWO2005033321), AAVhu.4 (SEQ ID NO: 141 of WO2005033321), AAVhu.40 (SEQID NO: 87 of WO2005033321), AAVhu.41 (SEQ ID NO: 91 of WO2005033321),AAVhu.42 (SEQ ID NO: 85 of WO2005033321), AAVhu.43 (SEQ ID NO: 160 ofWO2005033321), AAVhu.44 (SEQ ID NO: 144 of WO2005033321), AAVhu.45 (SEQID NO: 127 of WO2005033321), AAVhu.46 (SEQ ID NO: 159 of WO2005033321),AAVhu.47 (SEQ ID NO: 128 of WO2005033321), AAVhu.48 (SEQ ID NO: 157 ofWO2005033321), AAVhu.49 (SEQ ID NO: 189 of WO2005033321), AAVhu.51 (SEQID NO: 190 of WO2005033321), AAVhu.52 (SEQ ID NO: 191 of WO2005033321),AAVhu.53 (SEQ ID NO: 186 of WO2005033321), AAVhu.54 (SEQ ID NO: 188 ofWO2005033321), AAVhu.55 (SEQ ID NO: 187 of WO2005033321), AAVhu.56 (SEQID NO: 192 of WO2005033321), AAVhu.57 (SEQ ID NO: 193 of WO2005033321),AAVhu.58 (SEQ ID NO: 194 of WO2005033321), AAVhu.6 (SEQ ID NO: 84 ofWO2005033321), AAVhu.60 (SEQ ID NO: 184 of WO2005033321), AAVhu.61 (SEQID NO: 185 of WO2005033321), AAVhu.63 (SEQ ID NO: 195 of WO2005033321),AAVhu.64 (SEQ ID NO: 196 of WO2005033321), AAVhu.66 (SEQ ID NO: 197 ofWO2005033321), AAVhu.67 (SEQ ID NO: 198 of WO2005033321), AAVhu.7 (SEQID NO: 150 of WO2005033321), AAVhu.8 (WO2005033321 SEQ ID NO: 12),AAVhu.9 (SEQ ID NO: 155 of WO2005033321), AAVLG-10/rh.40 (SEQ ID No: 14of WO2005033321), AAVLG-4/rh.38 (SEQ ID NO: 86 of WO2005033321),AAVLG-4/rh.38 (SEQ ID No: 7 of WO2005033321), AAVN721-8/rh.43 (SEQ IDNO: 163 of WO2005033321), AAVN721-8/rh.43 (SEQ ID No: 43 ofWO2005033321), AAVpi.1 (WO2005033321 SEQ ID NO: 28), AAVpi.2(WO2005033321 SEQ ID NO: 30), AAVpi.3 (WO2005033321 SEQ ID NO: 29),AAVrh.38 (SEQ ID NO: 86 of WO2005033321), AAVrh.40 (SEQ ID NO: 92 ofWO2005033321), AAVrh.43 (SEQ ID NO: 163 of WO2005033321), AAVrh.44(WO2005033321 SEQ ID NO: 34), AAVrh.45 (WO2005033321 SEQ ID NO: 41),AAVrh.47 (WO2005033321 SEQ ID NO: 38), AAVrh.48 (SEQ ID NO: 115 ofWO2005033321), AAVrh.49 (SEQ ID NO: 103 of WO2005033321), AAVrh.50 (SEQID NO: 108 of WO2005033321), AAVrh.51 (SEQ ID NO: 104 of WO2005033321),AAVrh.52 (SEQ ID NO: 96 of WO2005033321), AAVrh.53 (SEQ ID NO: 97 ofWO2005033321), AAVrh.55 (WO2005033321 SEQ ID NO: 37), AAVrh.56 (SEQ IDNO: 152 of WO2005033321), AAVrh.57 (SEQ ID NO: 105 of WO2005033321),AAVrh.58 (SEQ ID NO: 106 of WO2005033321), AAVrh.59 (WO2005033321 SEQ IDNO: 42), AAVrh.60 (WO2005033321 SEQ ID NO: 31), AAVrh.61 (SEQ ID NO: 107of WO2005033321), AAVrh.62 (SEQ ID NO: 114 of WO2005033321), AAVrh.64(SEQ ID NO: 99 of WO2005033321), AAVrh.65 (WO2005033321 SEQ ID NO: 35),AAVrh.68 (WO2005033321 SEQ ID NO: 16), AAVrh.69 (WO2005033321 SEQ ID NO:39), AAVrh.70 (WO2005033321 SEQ ID NO: 20), AAVrh.72 (WO2005033321 SEQID NO: 9), or variants thereof including, but not limited to, AAVcy.2,AAVcy.3, AAVcy.4, AAVcy.5, AAVcy.6, AAVrh.12, AAVrh.17, AAVrh.18,AAVrh.19, AAVrh.21, AAVrh.22, AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.25/4215, AAVrh.31, AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35, AAVrh.36,AAVrh.37, AAVrh14. Non limiting examples of variants include SEQ ID NO:13, 15, 17, 19, 24, 36, 40, 45, 47, 48, 51-54, 60-62, 64-77, 79, 80, 82,89, 90, 93-95, 98, 100, 101, 109-113, 118-120, 124, 126, 131, 139, 142,151, 154, 158, 161, 162, 165-183, 202, 204-212, 215, 219, 224-236, ofWO2005033321, the contents of which are herein incorporated by referencein their entirety.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in International Publication No. WO2015168666, the contents ofwhich are herein incorporated by reference in their entirety, such as,but not limited to, AAVrh8R (SEQ ID NO: 9 of WO2015168666), AAVrh8RA586R mutant (SEQ ID NO: 10 of WO2015168666), AAVrh8R R533A mutant (SEQID NO: 11 of WO2015168666), or variants thereof.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in U.S. Pat. No. 9,233,131, the contents of which are hereinincorporated by reference in their entirety, such as, but not limitedto, AAVhE1.1 (SEQ ID NO:44 of U.S. Pat. No. 9,233,131), AAVhEr1.5 (SEQID NO:45 of U.S. Pat. No. 9,233,131), AAVhER1.14 (SEQ ID NO:46 of U.S.Pat. No. 9,233,131), AAVhEr1.8 (SEQ ID NO:47 of U.S. Pat. No.9,233,131), AAVhEr1.16 (SEQ ID NO:48 of U.S. Pat. No. 9,233,131),AAVhEr1.18 (SEQ ID NO:49 of U.S. Pat. No. 9,233,131), AAVhEr1.35 (SEQ IDNO:50 of U.S. Pat. No. 9,233,131), AAVhEr1.7 (SEQ ID NO:51 of U.S. Pat.No. 9,233,131), AAVhEr1.36 (SEQ ID NO:52 of U.S. Pat. No. 9,233,131),AAVhEr2.29 (SEQ ID NO:53 of U.S. Pat. No. 9,233,131), AAVhEr2.4 (SEQ IDNO:54 of U.S. Pat. No. 9,233,131), AAVhEr2.16 (SEQ ID NO:55 of U.S. Pat.No. 9,233,131), AAVhEr2.30 (SEQ ID NO:56 of U.S. Pat. No. 9,233,131),AAVhEr2.31 (SEQ ID NO:58 of U.S. Pat. No. 9,233,131), AAVhEr2.36 (SEQ IDNO:57 of U.S. Pat. No. 9,233,131), AAVhER1.23 (SEQ ID NO:53 of U.S. Pat.No. 9,233,131), AAVhEr3.1 (SEQ ID NO:59 of U.S. Pat. No. 9,233,131),AAV2.5T (SEQ ID NO:42 of U.S. Pat. No. 9,233,131), or variants thereof.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in United States Patent Publication No. US20150376607, thecontents of which are herein incorporated by reference in theirentirety, such as, but not limited to, AAV-PAEC (SEQ ID NO:1 ofUS20150376607), AAV-LK01 (SEQ ID NO:2 of US20150376607), AAV-LK02 (SEQID NO:3 of US20150376607), AAV-LK03 (SEQ ID NO:4 of US20150376607),AAV-LK04 (SEQ ID NO:5 of US20150376607), AAV-LK05 (SEQ ID NO:6 ofUS20150376607), AAV-LK06 (SEQ ID NO:7 of US20150376607), AAV-LK07 (SEQID NO:8 of US20150376607), AAV-LK08 (SEQ ID NO:9 of US20150376607),AAV-LK09 (SEQ ID NO:10 of US20150376607), AAV-LK10 (SEQ ID NO:11 ofUS20150376607), AAV-LK11 (SEQ ID NO:12 of US20150376607), AAV-LK12 (SEQID NO:13 of US20150376607), AAV-LK13 (SEQ ID NO:14 of US20150376607),AAV-LK14 (SEQ ID NO:15 of US20150376607), AAV-LK15 (SEQ ID NO:16 ofUS20150376607), AAV-LK16 (SEQ ID NO:17 of US20150376607), AAV-LK17 (SEQID NO:18 of US20150376607), AAV-LK18 (SEQ ID NO:19 of US20150376607),AAV-LK19 (SEQ ID NO:20 of US20150376607), AAV-PAEC2 (SEQ ID NO:21 ofUS20150376607), AAV-PAEC4 (SEQ ID NO:22 of US20150376607), AAV-PAEC6(SEQ ID NO:23 of US20150376607), AAV-PAEC7 (SEQ ID NO:24 ofUS20150376607), AAV-PAEC8 (SEQ ID NO:25 of US20150376607), AAV-PAEC11(SEQ ID NO:26 of US20150376607), AAV-PAEC12 (SEQ ID NO:27, ofUS20150376607), or variants thereof.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in U.S. Pat. No. 9,163,261, the contents of which are hereinincorporated by reference in their entirety, such as, but not limitedto, AAV-2-pre-miRNA-101 (SEQ ID NO: 1 U.S. Pat. No. 9,163,261), orvariants thereof.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in United States Patent Publication No. US20150376240, thecontents of which are herein incorporated by reference in theirentirety, such as, but not limited to, AAV-8h (SEQ ID NO: 6 ofUS20150376240), AAV-8b (SEQ ID NO: 5 of US20150376240), AAV-h (SEQ IDNO: 2 of US20150376240), AAV-b (SEQ ID NO: 1 of US20150376240), orvariants thereof.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in United States Patent Publication No. US20160017295, thecontents of which are herein incorporated by reference in theirentirety, such as, but not limited to, AAV SM 10-2 (SEQ ID NO: 22 ofUS20160017295), AAV Shuffle 100-1 (SEQ ID NO: 23 of US20160017295), AAVShuffle 100-3 (SEQ ID NO: 24 of US20160017295), AAV Shuffle 100-7 (SEQID NO: 25 of US20160017295), AAV Shuffle 10-2 (SEQ ID NO: 34 ofUS20160017295), AAV Shuffle 10-6 (SEQ ID NO: 35 of US20160017295), AAVShuffle 10-8 (SEQ ID NO: 36 of US20160017295), AAV Shuffle 100-2 (SEQ IDNO: 37 of US20160017295), AAV SM 10-1 (SEQ ID NO: 38 of US20160017295),AAV SM 10-8 (SEQ ID NO: 39 of US20160017295), AAV SM 100-3 (SEQ ID NO:40 of US20160017295), AAV SM 100-10 (SEQ ID NO: 41 of US20160017295), orvariants thereof.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in United States Patent Publication No. US20150238550, thecontents of which are herein incorporated by reference in theirentirety, such as, but not limited to, BNP61 AAV (SEQ ID NO: 1 ofUS20150238550), BNP62 AAV (SEQ ID NO: 3 of US20150238550), BNP63 AAV(SEQ ID NO: 4 of US20150238550), or variants thereof.

In some embodiments, the AAV serotype may be or may have a sequence asdescribed in United States Patent Publication No. US20150315612, thecontents of which are herein incorporated by reference in theirentirety, such as, but not limited to, AAVrh.50 (SEQ ID NO: 108 ofUS20150315612), AAVrh.43 (SEQ ID NO: 163 of US20150315612), AAVrh.62(SEQ ID NO: 114 of US20150315612), AAVrh.48 (SEQ ID NO: 115 ofUS20150315612), AAVhu.19 (SEQ ID NO: 133 of US20150315612), AAVhu.11(SEQ ID NO: 153 of US20150315612), AAVhu.53 (SEQ ID NO: 186 ofUS20150315612), AAV4-8/rh.64 (SEQ ID No: 15 ofUS20150315612),AAVLG-9/hu.39 (SEQ ID No: 24 of US20150315612),AAV54.5/hu.23 (SEQ ID No: 60 of US20150315612), AAV54.2/hu.22 (SEQ IDNo: 67 of US20150315612), AAV54.7/hu.24 (SEQ ID No: 66 ofUS20150315612), AAV54.1/hu.21 (SEQ ID No: 65 of US20150315612),AAV54.4R/hu.27 (SEQ ID No: 64 of US20150315612), AAV46.2/hu.28 (SEQ IDNo: 68 of US20150315612), AAV46.6/hu.29 (SEQ ID No: 69 ofUS20150315612), AAV128.1/hu.43 (SEQ ID No: 80 of US20150315612), orvariants thereof.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in International Publication No. WO2015121501, the contents ofwhich are herein incorporated by reference in their entirety, such as,but not limited to, true type AAV (ttAAV) (SEQ ID NO: 2 ofWO2015121501), “UPenn AAV10” (SEQ ID NO: 8 of WO2015121501), “JapaneseAAV10” (SEQ ID NO: 9 of WO2015121501), or variants thereof.

According to the present disclosure, AAV capsid serotype selection oruse may be from a variety of species. In one embodiment, the AAV may bean avian AAV (AAAV). The AAAV serotype may be, or have, a sequence asdescribed in U.S. Pat. No. 9,238,800, the contents of which are hereinincorporated by reference in their entirety, such as, but not limitedto, AAAV (SEQ ID NO: 1, 2, 4, 6, 8, 10, 12, and 14 of U.S. Pat. No.9,238,800), or variants thereof.

In one embodiment, the AAV may be a bovine AAV (BAAV). The BAAV serotypemay be, or have, a sequence as described in U.S. Pat. No. 9,193,769, thecontents of which are herein incorporated by reference in theirentirety, such as, but not limited to, BAAV (SEQ ID NO: 1 and 6 of U.S.Pat. No. 9,193,769), or variants thereof. The BAAV serotype may be orhave a sequence as described in U.S. Pat. No. 7,427,396, the contents ofwhich are herein incorporated by reference in their entirety, such as,but not limited to, BAAV (SEQ ID NO: 5 and 6 of U.S. Pat. No.7,427,396), or variants thereof.

In one embodiment, the AAV may be a caprine AAV. The caprine AAVserotype may be, or have, a sequence as described in U.S. Pat. No.7,427,396, the contents of which are herein incorporated by reference intheir entirety, such as, but not limited to, caprine AAV (SEQ ID NO: 3of U.S. Pat. No. 7,427,396), or variants thereof.

In other embodiments the AAV may be engineered as a hybrid AAV from twoor more parental serotypes. In one embodiment, the AAV may be AAV2G9which comprises sequences from AAV2 and AAV9. The AAV2G9 AAV serotypemay be, or have, a sequence as described in United States PatentPublication No. US20160017005, the contents of which are hereinincorporated by reference in its entirety.

In one embodiment, the AAV may be a serotype generated by the AAV9capsid library with mutations in amino acids 390-627 (VP1 numbering) asdescribed by Pulicherla et al. (Molecular Therapy 19(6):1070-1078(2011), the contents of which are herein incorporated by reference intheir entirety. The serotype and corresponding nucleotide and amino acidsubstitutions may be, but is not limited to, AAV9.1 (G1594C; D532H),AAV6.2 (T1418A and T1436X; V473D and I479K), AAV9.3 (T1238A; F413Y),AAV9.4 (T1250C and A1617T; F417S), AAV9.5 (A1235G, A1314T, A1642G,C1760T; Q412R, T548A, A587V), AAV9.6 (T1231A; F411I), AAV9.9 (G1203A,G1785T; W595C), AAV9.10 (A1500G, T1676C; M559T), AAV9.11 (A1425T,A1702C, A1769T; T568P, Q590L), AAV9.13 (A1369C, A1720T; N457H, T574S),AAV9.14 (T1340A, T1362C, T1560C, G1713A; L447H), AAV9.16 (A1775T;Q592L), AAV9.24 (T1507C, T1521G; W503R), AAV9.26 (A1337G, A1769C; Y446C,Q590P), AAV9.33 (A1667C; D556A), AAV9.34 (A1534G, C1794T; N512D),AAV9.35 (A1289T, T1450A, C1494T, A1515T, C1794A, G1816A; Q430L, Y484N,N98K, V6061), AAV9.40 (A1694T, E565V), AAV9.41 (A1348T, T1362C; T450S),AAV9.44 (A1684C, A1701T, A1737G; N562H, K567N), AAV9.45 (A1492T, C1804T;N498Y, L602F), AAV9.46 (G1441C, T1525C, T1549G; G481R, W509R, L517V),9.47 (G1241A, G1358A, A1669G, C1745T; S414N, G453D, K557E, T582I),AAV9.48 (C1445T, A1736T; P482L, Q579L), AAV9.50 (A1638T, C1683T, T1805A;Q546H, L602H), AAV9.53 (G1301A, A1405C, C1664T, G1811T; R134Q, S469R,A555V, G604V), AAV9.54 (C1531A, T1609A; L51I, L537M), AAV9.55 (T1605A;F535L), AAV9.58 (C1475T, C1579A; T492I, H527N), AAV.59 (T1336C; Y446H),AAV9.61 (A1493T; N498I), AAV9.64 (C1531A, A1617T; L51I), AAV9.65(C1335T, T1530C, C1568A; A523D), AAV9.68 (C1510A; P504T), AAV9.80(G1441A,G481R), AAV9.83 (C1402A, A1500T; P468T, E500D), AAV9.87 (T1464C,T1468C; S490P), AAV9.90 (A1196T; Y399F), AAV9.91 (T1316G, A1583T,C1782G, T1806C; L439R, K528I), AAV9.93 (A1273G, A1421G, A1638C, C1712T,G1732A, A1744T, A1832T; S425G, Q474R, Q546H, P571L, G578R, T582S,D611V), AAV9.94 (A1675T; M559L) and AAV9.95 (T1605A; F535L).

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in International Publication No. WO2016049230, the contents ofwhich are herein incorporated by reference in their entirety, such as,but not limited to AAVF1/HSC1 (SEQ ID NO: 2 and 20 of WO2016049230),AAVF2/HSC2 (SEQ ID NO: 3 and 21 of WO2016049230), AAVF3/HSC3 (SEQ ID NO:5 and 22 of WO2016049230), AAVF4/HSC4 (SEQ ID NO: 6 and 23 ofWO2016049230), AAVF5/HSC5 (SEQ ID NO: 11 and 25 of WO2016049230),AAVF6/HSC6 (SEQ ID NO: 7 and 24 of WO2016049230), AAVF7/HSC7 (SEQ ID NO:8 and 27 of WO2016049230), AAVF8/HSC8 (SEQ ID NO: 9 and 28 ofWO2016049230), AAVF9/HSC9 (SEQ ID NO: 10 and 29 of WO2016049230),AAVF11/HSC11 (SEQ ID NO: 4 and 26 of WO2016049230), AAVF12/HSC12 (SEQ IDNO: 12 and 30 of WO2016049230), AAVF13/HSC13 (SEQ ID NO: 14 and 31 ofWO2016049230), AAVF14/HSC14 (SEQ ID NO: 15 and 32 of WO2016049230),AAVF15/HSC15 (SEQ ID NO: 16 and 33 of WO2016049230), AAVF16/HSC16 (SEQID NO: 17 and 34 of WO2016049230), AAVF17/HSC17 (SEQ ID NO: 13 and 35 ofWO2016049230), or variants or derivatives thereof.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in U.S. Pat. No. 8,734,809, the contents of which are hereinincorporated by reference in their entirety, such as, but not limitedto, AAV CBr-E1 (SEQ ID NO: 13 and 87 of U.S. Pat. No. 8,734,809), AAVCBr-E2 (SEQ ID NO: 14 and 88 of U.S. Pat. No. 8,734,809), AAV CBr-E3(SEQ ID NO: 15 and 89 of U.S. Pat. No. 8,734,809), AAV CBr-E4 (SEQ IDNO: 16 and 90 of U.S. Pat. No. 8,734,809), AAV CBr-E5 (SEQ ID NO: 17 and91 of U.S. Pat. No. 8,734,809), AAV CBr-e5 (SEQ ID NO: 18 and 92 of U.S.Pat. No. 8,734,809), AAV CBr-E6 (SEQ ID NO: 19 and 93 of U.S. Pat. No.8,734,809), AAV CBr-E7 (SEQ ID NO: 20 and 94 of U.S. Pat. No.8,734,809), AAV CBr-E8 (SEQ ID NO: 21 and 95 of U.S. Pat. No.8,734,809), AAV CLv-D1 (SEQ ID NO: 22 and 96 of U.S. Pat. No.8,734,809), AAV CLv-D2 (SEQ ID NO: 23 and 97 of U.S. Pat. No.8,734,809), AAV CLv-D3 (SEQ ID NO: 24 and 98 of U.S. Pat. No.8,734,809), AAV CLv-D4 (SEQ ID NO: 25 and 99 of U.S. Pat. No.8,734,809), AAV CLv-D5 (SEQ ID NO: 26 and 100 of U.S. Pat. No.8,734,809), AAV CLv-D6 (SEQ ID NO: 27 and 101 of U.S. Pat. No.8,734,809), AAV CLv-D7 (SEQ ID NO: 28 and 102 of U.S. Pat. No.8,734,809), AAV CLv-D8 (SEQ ID NO: 29 and 103 of U.S. Pat. No.8,734,809), AAV CLv-E1 (SEQ ID NO: 13 and 87 of U.S. Pat. No.8,734,809), AAV CLv-R1 (SEQ ID NO: 30 and 104 of U.S. Pat. No.8,734,809), AAV CLv-R2 (SEQ ID NO: 31 and 105 of U.S. Pat. No.8,734,809), AAV CLv-R3 (SEQ ID NO: 32 and 106 of U.S. Pat. No.8,734,809), AAV CLv-R4 (SEQ ID NO: 33 and 107 of U.S. Pat. No.8,734,809), AAV CLv-R5 (SEQ ID NO: 34 and 108 of U.S. Pat. No.8,734,809), AAV CLv-R6 (SEQ ID NO: 35 and 109 of U.S. Pat. No.8,734,809), AAV CLv-R7 (SEQ ID NO: 36 and 110 of U.S. Pat. No.8,734,809), AAV CLv-R8 (SEQ ID NO: X and X of U.S. Pat. No. 8,734,809),AAV CLv-R9 (SEQ ID NO: X and X of U.S. Pat. No. 8,734,809), AAV CLg-F1(SEQ ID NO: 39 and 113 of U.S. Pat. No. 8,734,809), AAV CLg-F2 (SEQ IDNO: 40 and 114 of U.S. Pat. No. 8,734,809), AAV CLg-F3 (SEQ ID NO: 41and 115 of U.S. Pat. No. 8,734,809), AAV CLg-F4 (SEQ ID NO: 42 and 116of U.S. Pat. No. 8,734,809), AAV CLg-F5 (SEQ ID NO: 43 and 117 of U.S.Pat. No. 8,734,809), AAV CLg-F6 (SEQ ID NO: 43 and 117 of U.S. Pat. No.8,734,809), AAV CLg-F7 (SEQ ID NO: 44 and 118 of U.S. Pat. No.8,734,809), AAV CLg-F8 (SEQ ID NO: 43 and 117 of U.S. Pat. No.8,734,809), AAV CSp-1 (SEQ ID NO: 45 and 119 of U.S. Pat. No.8,734,809), AAV CSp-10 (SEQ ID NO: 46 and 120 of U.S. Pat. No.8,734,809), AAV CSp-11 (SEQ ID NO: 47 and 121 of U.S. Pat. No.8,734,809), AAV CSp-2 (SEQ ID NO: 48 and 122 of U.S. Pat. No.8,734,809), AAV CSp-3 (SEQ ID NO: 49 and 123 of U.S. Pat. No.8,734,809), AAV CSp-4 (SEQ ID NO: 50 and 124 of U.S. Pat. No.8,734,809), AAV CSp-6 (SEQ ID NO: 51 and 125 of U.S. Pat. No.8,734,809), AAV CSp-7 (SEQ ID NO: 52 and 126 of U.S. Pat. No.8,734,809), AAV CSp-8 (SEQ ID NO: 53 and 127 of U.S. Pat. No.8,734,809), AAV CSp-9 (SEQ ID NO: 54 and 128 of U.S. Pat. No.8,734,809), AAV CHt-2 (SEQ ID NO: 55 and 129 of U.S. Pat. No.8,734,809), AAV CHt-3 (SEQ ID NO: 56 and 130 of U.S. Pat. No.8,734,809), AAV CKd-1 (SEQ ID NO: 57 and 131 of U.S. Pat. No.8,734,809), AAV CKd-10 (SEQ ID NO: 58 and 132 of U.S. Pat. No.8,734,809), AAV CKd-2 (SEQ ID NO: 59 and 133 of U.S. Pat. No.8,734,809), AAV CKd-3 (SEQ ID NO: 60 and 134 of U.S. Pat. No.8,734,809), AAV CKd-4 (SEQ ID NO: 61 and 135 of U.S. Pat. No.8,734,809), AAV CKd-6 (SEQ ID NO: 62 and 136 of U.S. Pat. No.8,734,809), AAV CKd-7 (SEQ ID NO: 63 and 137 of U.S. Pat. No.8,734,809), AAV CKd-8 (SEQ ID NO: 64 and 138 of U.S. Pat. No.8,734,809), AAV CLv-1 (SEQ ID NO: 35 and 139 of U.S. Pat. No.8,734,809), AAV CLv-12 (SEQ ID NO: 66 and 140 of U.S. Pat. No.8,734,809), AAV CLv-13 (SEQ ID NO: 67 and 141 of U.S. Pat. No.8,734,809), AAV CLv-2 (SEQ ID NO: 68 and 142 of U.S. Pat. No.8,734,809), AAV CLv-3 (SEQ ID NO: 69 and 143 of U.S. Pat. No.8,734,809), AAV CLv-4 (SEQ ID NO: 70 and 144 of U.S. Pat. No.8,734,809), AAV CLv-6 (SEQ ID NO: 71 and 145 of U.S. Pat. No.8,734,809), AAV CLv-8 (SEQ ID NO: 72 and 146 of U.S. Pat. No.8,734,809), AAV CKd-B1 (SEQ ID NO: 73 and 147 of U.S. Pat. No.8,734,809), AAV CKd-B2 (SEQ ID NO: 74 and 148 of U.S. Pat. No.8,734,809), AAV CKd-B3 (SEQ ID NO: 75 and 149 of U.S. Pat. No.8,734,809), AAV CKd-B4 (SEQ ID NO: 76 and 150 of U.S. Pat. No.8,734,809), AAV CKd-B5 (SEQ ID NO: 77 and 151 of U.S. Pat. No.8,734,809), AAV CKd-B6 (SEQ ID NO: 78 and 152 of U.S. Pat. No.8,734,809), AAV CKd-B7 (SEQ ID NO: 79 and 153 of U.S. Pat. No.8,734,809), AAV CKd-B8 (SEQ ID NO: 80 and 154 of U.S. Pat. No.8,734,809), AAV CKd-H1 (SEQ ID NO: 81 and 155 of U.S. Pat. No.8,734,809), AAV CKd-H2 (SEQ ID NO: 82 and 156 of U.S. Pat. No.8,734,809), AAV CKd-H3 (SEQ ID NO: 83 and 157 of U.S. Pat. No.8,734,809), AAV CKd-H4 (SEQ ID NO: 84 and 158 of U.S. Pat. No.8,734,809), AAV CKd-H5 (SEQ ID NO: 85 and 159 of U.S. Pat. No.8,734,809), AAV CKd-H6 (SEQ ID NO: 77 and 151 of U.S. Pat. No.8,734,809), AAV CHt-1 (SEQ ID NO: 86 and 160 of U.S. Pat. No.8,734,809), AAV CLv1-1 (SEQ ID NO: 171 of U.S. Pat. No. 8,734,809), AAVCLv1-2 (SEQ ID NO: 172 of U.S. Pat. No. 8,734,809), AAV CLv1-3 (SEQ IDNO: 173 of U.S. Pat. No. 8,734,809), AAV CLv1-4 (SEQ ID NO: 174 of U.S.Pat. No. 8,734,809), AAV Clv1-7 (SEQ ID NO: 175 of U.S. Pat. No.8,734,809), AAV Clv1-8 (SEQ ID NO: 176 of U.S. Pat. No. 8,734,809), AAVClv1-9 (SEQ ID NO: 177 of US8734809), AAV Clv1-10 (SEQ ID NO: 178 ofU.S. Pat. No. 8,734,809), AAV.VR-355 (SEQ ID NO: 181 of U.S. Pat. No.8,734,809), AAV.hu.48R3 (SEQ ID NO: 183 of U.S. Pat. No. 8,734,809), orvariants or derivatives thereof.

In some embodiments, the AAV serotype may be, or have, a sequence asdescribed in International Publication No. WO2016065001, the contents ofwhich are herein incorporated by reference in their entirety, such as,but not limited to AAV CHt-P2 (SEQ ID NO: 1 and 51 of WO2016065001), AAVCHt-P5 (SEQ ID NO: 2 and 52 of WO2016065001), AAV CHt-P9 (SEQ ID NO: 3and 53 of WO2016065001), AAV CBr-7.1 (SEQ ID NO: 4 and 54 ofWO2016065001), AAV CBr-7.2 (SEQ ID NO: 5 and 55 of WO2016065001), AAVCBr-7.3 (SEQ ID NO: 6 and 56 of WO2016065001), AAV CBr-7.4 (SEQ ID NO: 7and 57 of WO2016065001), AAV CBr-7.5 (SEQ ID NO: 8 and 58 ofWO2016065001), AAV CBr-7.7 (SEQ ID NO: 9 and 59 of WO2016065001), AAVCBr-7.8 (SEQ ID NO: 10 and 60 of WO2016065001), AAV CBr-7.10 (SEQ ID NO:11 and 61 of WO2016065001), AAV CKd-N3 (SEQ ID NO: 12 and 62 ofWO2016065001), AAV CKd-N4 (SEQ ID NO: 13 and 63 of WO2016065001), AAVCKd-N9 (SEQ ID NO: 14 and 64 of WO2016065001), AAV CLv-L4 (SEQ ID NO: 15and 65 of WO2016065001), AAV CLv-L5 (SEQ ID NO: 16 and 66 ofWO2016065001), AAV CLv-L6 (SEQ ID NO: 17 and 67 of WO2016065001), AAVCLv-K1 (SEQ ID NO: 18 and 68 of WO2016065001), AAV CLv-K3 (SEQ ID NO: 19and 69 of WO2016065001), AAV CLv-K6 (SEQ ID NO: 20 and 70 ofWO2016065001), AAV CLv-M1 (SEQ ID NO: 21 and 71 of WO2016065001), AAVCLv-M11 (SEQ ID NO: 22 and 72 of WO2016065001), AAV CLv-M2 (SEQ ID NO:23 and 73 of WO2016065001), AAV CLv-M5 (SEQ ID NO: 24 and 74 ofWO2016065001), AAV CLv-M6 (SEQ ID NO: 25 and 75 of WO2016065001), AAVCLv-M7 (SEQ ID NO: 26 and 76 of WO2016065001), AAV CLv-M8 (SEQ ID NO: 27and 77 of WO2016065001), AAV CLv-M9 (SEQ ID NO: 28 and 78 ofWO2016065001), AAV CHt-P1 (SEQ ID NO: 29 and 79 of WO2016065001), AAVCHt-P6 (SEQ ID NO: 30 and 80 of WO2016065001), AAV CHt-P8 (SEQ ID NO: 31and 81 of WO2016065001), AAV CHt-6.1 (SEQ ID NO: 32 and 82 ofWO2016065001), AAV CHt-6.10 (SEQ ID NO: 33 and 83 of WO2016065001), AAVCHt-6.5 (SEQ ID NO: 34 and 84 of WO2016065001), AAV CHt-6.6 (SEQ ID NO:35 and 85 of WO2016065001), AAV CHt-6.7 (SEQ ID NO: 36 and 86 ofWO2016065001), AAV CHt-6.8 (SEQ ID NO: 37 and 87 of WO2016065001), AAVCSp-8.10 (SEQ ID NO: 38 and 88 of WO2016065001), AAV CSp-8.2 (SEQ ID NO:39 and 89 of WO2016065001), AAV CSp-8.4 (SEQ ID NO: 40 and 90 ofWO2016065001), AAV CSp-8.5 (SEQ ID NO: 41 and 91 of WO2016065001), AAVCSp-8.6 (SEQ ID NO: 42 and 92 of WO2016065001), AAV CSp-8.7 (SEQ ID NO:43 and 93 of WO2016065001), AAV CSp-8.8 (SEQ ID NO: 44 and 94 ofWO2016065001), AAV CSp-8.9 (SEQ ID NO: 45 and 95 of WO2016065001), AAVCBr-B7.3 (SEQ ID NO: 46 and 96 of WO2016065001), AAV CBr-B7.4 (SEQ IDNO: 47 and 97 of WO2016065001), AAV3B (SEQ ID NO: 48 and 98 ofWO2016065001), AAV4 (SEQ ID NO: 49 and 99 of WO2016065001), AAV5 (SEQ IDNO: 50 and 100 of WO2016065001), or variants or derivatives thereof.

In some embodiments, the AAV serotype may be, or have, a modification asdescribed in United States Publication No. US 20160361439, the contentsof which are herein incorporated by reference in their entirety, such asbut not limited to, Y252F, Y272F, Y444F, Y500F, Y700F, Y704F, Y730F,Y275F, Y281F, Y508F, Y576F, Y612G, Y673F, and Y720F of the wild-typeAAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11,AAV12,and hybrids thereof.

In some embodiments, the AAV serotype may be, or have, a mutation asdescribed in U.S. Pat. No. 9,546,112, the contents of which are hereinincorporated by reference in their entirety, such as, but not limitedto, at least two, but not all the F129L, D418E, K531E, L584F, V598A andH642N mutations in the sequence of AAV6 (SEQ ID NO:4 of U.S. Pat. No.9,546,112), AAV1 (SEQ ID NO:6 of U.S. Pat. No. 9,546,112), AAV2, AAV3,AAV4, AAV5, AAV7, AAV9, AAV10 or AAV11 or derivatives thereof. In yetanother embodiment, the AAV serotype may be, or have, an AAV6 sequencecomprising the K531E mutation (SEQ ID NO:5 of U.S. Pat. No. 9,546,112).

In some embodiments, the AAV serotype may be, or have, a mutation in theAAV1 sequence, as described in in United States Publication No. US20130224836, the contents of which are herein incorporated by referencein their entirety, such as, but not limited to, at least one of thesurface-exposed tyrosine residues, preferably, at positions 252, 273,445, 701, 705 and 731 of AAV1 (SEQ ID NO: 2 of US 20130224836)substituted with another amino acid, preferably with a phenylalanineresidue. In one embodiment, the AAV serotype may be, or have, a mutationin the AAV9 sequence, such as, but not limited to, at least one of thesurface-exposed tyrosine residues, preferably, at positions 252, 272,444, 500, 700, 704 and 730 of AAV2 (SEQ ID NO: 4 of US 20130224836)substituted with another amino acid, preferably with a phenylalanineresidue. In one embodiment, the tyrosine residue at position 446 of AAV9(SEQ ID NO: 6 US 20130224836) is substituted with a phenylalanineresidue.

In some embodiments, the serotype may be AAV2 or a variant thereof, asdescribed in International Publication No. WO2016130589, hereinincorporated by reference in its entirety. The amino acid sequence ofAAV2 may comprise N587A, E548A, or N708A mutations. In one embodiment,the amino acid sequence of any AAV may comprise a V708K mutation.

In one embodiment, the AAV may be a serotype selected from any of thosefound in Table 1.

In one embodiment, the AAV may comprise a sequence, fragment or variantthereof, of the sequences in Table 1.

In one embodiment, the AAV may be encoded by a sequence, fragment orvariant as described in Table 1.

TABLE 1 AAV Serotypes Serotype SEQ ID NO Reference Information AAVPHP.Bor G2B-26 1 WO2015038958 SEQ ID NO: 8 and 13 AAVPHP.B 2 WO2015038958 SEQID NO: 9 AAVG2B-13 3 WO2015038958 SEQ ID NO: 12 AAVTH1.1-32 4WO2015038958 SEQ ID NO: 14 AAVTH1.1-35 5 WO2015038958 SEQ ID NO: 15 AAV16 US20150159173 SEQ ID NO: 11, US20150315612 SEQ ID NO: 202 AAV1 7US20160017295 SEQ ID NO: 1US20030138772 SEQ ID NO: 64, US20150159173 SEQID NO: 27, US20150315612 SEQ ID NO: 219, U.S. Pat. No. 7,198,951 SEQ IDNO: 5 AAV1 8 US20030138772 SEQ ID NO: 6 AAV1.3 9 US20030138772 SEQ IDNO: 14 AAV10 10 US20030138772 SEQ ID NO: 117 AAV10 11 WO2015121501 SEQID NO: 9 AAV10 12 WO2015121501 SEQ ID NO: 8 AAV11 13 US20030138772 SEQID NO: 118 AAV12 14 US20030138772 SEQ ID NO: 119 AAV2 15 US20150159173SEQ ID NO: 7, US20150315612 SEQ ID NO: 211 AAV2 16 US20030138772 SEQ IDNO: 70, US20150159173 SEQ ID NO: 23, US20150315612 SEQ ID NO: 221,US20160017295 SEQ ID NO: 2, U.S. Pat. No. 6,156,303 SEQ ID NO: 4, U.S.Pat. No. 7,198,951 SEQ ID NO: 4, WO2015121501 SEQ ID NO: 1 AAV2 17 U.S.Pat. No. 6,156,303 SEQ ID NO: 8 AAV2 18 US20030138772 SEQ ID NO: 7 AAV219 U.S. Pat. No. 6,156,303 SEQ ID NO: 3 AAV2.5T 20 U.S. Pat. No9,233,131 SEQ ID NO: 42 AAV223.10 21 US20030138772 SEQ ID NO: 75AAV223.2 22 US20030138772 SEQ ID NO: 49 AAV223.2 23 US20030138772 SEQ IDNO: 76 AAV223.4 24 US20030138772 SEQ ID NO: 50 AAV223.4 25 US20030138772SEQ ID NO: 73 AAV223.5 26 US20030138772 SEQ ID NO: 51 AAV223.5 27US20030138772 SEQ ID NO: 74 AAV223.6 28 US20030138772 SEQ ID NO: 52AAV223.6 29 US20030138772 SEQ ID NO: 78 AAV223.7 30 US20030138772 SEQ IDNO: 53 AAV223.7 31 US20030138772 SEQ ID NO: 77 AAV29.3 32 US20030138772SEQ ID NO: 82 AAV29.4 33 US20030138772 SEQ ID NO: 12 AAV29.5 34US20030138772 SEQ ID NO: 83 AAV29.5 (AAVbb.2) 35 US20030138772 SEQ IDNO: 13 AAV3 36 US20150159173 SEQ ID NO: 12 AAV3 37 US20030138772 SEQ IDNO: 71, US20150159173 SEQ ID NO: 28, US20160017295 SEQ ID NO: 3, U.S.Pat. No 7,198,951 SEQ ID NO: 6 AAV3 38 US20030138772 SEQ ID NO: 8AAV3.3b 39 US20030138772 SEQ ID NO: 72 AAV3-3 40 US20150315612 SEQ IDNO: 200 AAV3-3 41 US20150315612 SEQ ID NO: 217 AAV3a 42 U.S. Pat. No.6,156,303 SEQ ID NO: 5 AAV3a 43 U.S. Pat. No. 6,156,303 SEQ ID NO: 9AAV3b 44 U.S. Pat. No. 6,156,303 SEQ ID NO: 6 AAV3b 45 U.S. Pat. No.6,156,303 SEQ ID NO: 10 AAV3b 46 U.S. Pat. No. 6,156,303 SEQ ID NO: 1AAV4 47 US20140348794 SEQ ID NO: 17 AAV4 48 US20140348794 SEQ ID NO: 5AAV4 49 US20140348794 SEQ ID NO: 3 AAV4 50 US20140348794 SEQ ID NO: 14AAV4 51 US20140348794 SEQ ID NO: 15 AAV4 52 US20140348794 SEQ ID NO: 19AAV4 53 US20140348794 SEQ ID NO: 12 AAV4 54 US20140348794 SEQ ID NO: 13AAV4 55 US20140348794 SEQ ID NO: 7 AAV4 56 US20140348794 SEQ ID NO: 8AAV4 57 US20140348794 SEQ ID NO: 9 AAV4 58 US20140348794 SEQ ID NO: 2AAV4 59 US20140348794 SEQ ID NO: 10 AAV4 60 US20140348794 SEQ ID NO: 11AAV4 61 US20140348794 SEQ ID NO: 18 AAV4 62 US20030138772 SEQ ID NO: 63,US20160017295 SEQ ID NO: 4, US20140348794 SEQ ID NO: 4 AAV4 63US20140348794 SEQ ID NO: 16 AAV4 64 US20140348794 SEQ ID NO: 20 AAV4 65US20140348794 SEQ ID NO: 6 AAV4 66 US20140348794 SEQ ID NO: 1 AAV42.2 67US20030138772 SEQ ID NO: 9 AAV42.2 68 US20030138772 SEQ ID NO: 102AAV42.3b 69 US20030138772 SEQ ID NO: 36 AAV42.3B 70 US20030138772 SEQ IDNO: 107 AAV42.4 71 US20030138772 SEQ ID NO: 33 AAV42.4 72 US20030138772SEQ ID NO: 88 AAV42.8 73 US20030138772 SEQ ID NO: 27 AAV42.8 74US20030138772 SEQ ID NO: 85 AAV43.1 75 US20030138772 SEQ ID NO: 39AAV43.1 76 US20030138772 SEQ ID NO: 92 AAV43.12 77 US20030138772 SEQ IDNO: 41 AAV43.12 78 US20030138772 SEQ ID NO: 93 AAV43.20 79 US20030138772SEQ ID NO: 42 AAV43.20 80 US20030138772 SEQ ID NO: 99 AAV43.21 81US20030138772 SEQ ID NO: 43 AAV43.21 82 US20030138772 SEQ ID NO: 96AAV43.23 83 US20030138772 SEQ ID NO: 44 AAV43.23 84 US20030138772 SEQ IDNO: 98 AAV43.25 85 US20030138772 SEQ ID NO: 45 AAV43.25 86 US20030138772SEQ ID NO: 97 AAV43.5 87 US20030138772 SEQ ID NO: 40 AAV43.5 88US20030138772 SEQ ID NO: 94 AAV4-4 89 US20150315612 SEQ ID NO: 201AAV4-4 90 US20150315612 SEQ ID NO: 218 AAV44.1 91 US20030138772 SEQ IDNO: 46 AAV44.1 92 US20030138772 SEQ ID NO: 79 AAV44.5 93 US20030138772SEQ ID NO: 47 AAV44.5 94 US20030138772 SEQ ID NO: 80 AAV4407 95US20150315612 SEQ ID NO: 90 AAV5 96 U.S. Pat. No. 7,427,396 SEQ ID NO: 1AAV5 97 US20030138772 SEQ ID NO: 114 AAV5 98 US20160017295 SEQ ID NO: 5,U.S. Pat. No. 7,427,396 SEQ ID NO: 2, US20150315612 SEQ ID NO: 216 AAV599 US20150315612 SEQ ID NO: 199 AAV6 100 US20150159173 SEQ ID NO: 13AAV6 101 US20030138772 SEQ ID NO: 65, US20150159173 SEQ ID NO: 29,US20160017295 SEQ ID NO: 6, U.S. Pat. No. 6,156,303 SEQ ID NO: 7 AAV6102 U.S. Pat. No. 6,156,303 SEQ ID NO: 11 AAV6 103 U.S. Pat. No.6,156,303 SEQ ID NO: 2 AAV6 104 US20150315612 SEQ ID NO: 203 AAV6 105US20150315612 SEQ ID NO: 220 AAV6.1 106 US20150159173 AAV6.12 107US20150159173 AAV6.2 108 US20150159173 AAV7 109 US20150159173 SEQ ID NO:14 AAV7 110 US20150315612 SEQ ID NO: 183 AAV7 111 US20030138772 SEQ IDNO: 2, US20150159173 SEQ ID NO: 30, US20150315612 SEQ ID NO: 181,US20160017295 SEQ ID NO: 7 AAV7 112 US20030138772 SEQ ID NO: 3 AAV7 113US20030138772 SEQ ID NO: 1, US20150315612 SEQ ID NO: 180 AAV7 114US20150315612 SEQ ID NO: 213 AAV7 115 US20150315612 SEQ ID NO: 222 AAV8116 US20150159173 SEQ ID NO: 15 AAV8 117 US20150376240 SEQ ID NO: 7 AAV8118 US20030138772 SEQ ID NO: 4, US20150315612 SEQ ID NO: 182 AAV8 119US20030138772 SEQ ID NO: 95, US20140359799 SEQ ID NO: 1, US20150159173SEQ ID NO: 31, US20160017295 SEQ ID NO: 8, U.S. Pat. No. 7,198,951 SEQID NO: 7, US20150315612 SEQ ID NO: 223 AAV8 120 US20150376240 SEQ ID NO:8 AAV8 121 US20150315612 SEQ ID NO: 214 AAV-8b 122 US20150376240 SEQ IDNO: 5 AAV-8b 123 US20150376240 SEQ ID NO: 3 AAV-8h 124 US20150376240 SEQID NO: 6 AAV-8h 125 US20150376240 SEQ ID NO: 4 AAV9 126 US20030138772SEQ ID NO: 5 AAV9 127 U.S. Pat. No. 7,198,951 SEQ ID NO: 1 AAV9 128US20160017295 SEQ ID NO: 9 AAV9 129 US20030138772 SEQ ID NO: 100, U.S.Pat. No. 7,198,951 SEQ ID NO: 2 AAV9 130 U.S. Pat. No. 7,198,951 SEQ IDNO: 3 AAV9 (AAVhu.14) 131 U.S. Pat. No. 7,906,111 SEQ ID NO: 3;WO2015038958 SEQ ID NO: 11 AAV9 (AAVhu.14) 132 U.S. Pat. No. 7,906,111SEQ ID NO: 123; WO2015038958 SEQ ID NO: 2 AAVA3.1 133 US20030138772 SEQID NO: 120 AAVA3.3 134 US20030138772 SEQ ID NO: 57 AAVA3.3 135US20030138772 SEQ ID NO: 66 AAVA3.4 136 US20030138772 SEQ ID NO: 54AAVA3.4 137 US20030138772 SEQ ID NO: 68 AAVA3.5 138 US20030138772 SEQ IDNO: 55 AAVA3.5 139 US20030138772 SEQ ID NO: 69 AAVA3.7 140 US20030138772SEQ ID NO: 56 AAVA3.7 141 US20030138772 SEQ ID NO: 67 AAV29.3 (AAVbb.1)142 US20030138772 SEQ ID NO: 11 AAVC2 143 US20030138772 SEQ ID NO: 61AAVCh.5 144 US20150159173 SEQ ID NO: 46, US20150315612 SEQ ID NO: 234AAVcy.2 (AAV13.3) 145 US20030138772 SEQ ID NO: 15 AAV24.1 146US20030138772 SEQ ID NO: 101 AAVcy.3 (AAV24.1) 147 US20030138772 SEQ IDNO: 16 AAV27.3 148 US20030138772 SEQ ID NO: 104 AAVcy.4 (AAV27.3) 149US20030138772 SEQ ID NO: 17 AAVcy.5 150 US20150315612 SEQ ID NO: 227AAV7.2 151 US20030138772 SEQ ID NO: 103 AAVcy.5 (AAV7.2) 152US20030138772 SEQ ID NO: 18 AAV16.3 153 US20030138772 SEQ ID NO: 105AAVcy.6 (AAV16.3) 154 US20030138772 SEQ ID NO: 10 AAVcy.5 155US20150159173 SEQ ID NO: 8 AAVcy.5 156 US20150159173 SEQ ID NO: 24AAVCy.5R1 157 US20150159173 AAVCy.5R2 158 US20150159173 AAVCy.5R3 159US20150159173 AAVCy.5R4 160 US20150159173 AAVDJ 161 US20140359799 SEQ IDNO: 3, U.S. Pat. No. 7,588,772 SEQ ID NO: 2 AAVDJ 162 US20140359799 SEQID NO: 2, U.S. Pat. No. 7,588,772 SEQ ID NO: 1 AAVDJ-8 163 U.S. Pat. No.7,588,772; Grimm et al 2008 AAVDJ-8 164 U.S. Pat. No. 7,588,772; Grimmet al 2008 AAVF5 165 US20030138772 SEQ ID NO: 110 AAVH2 166US20030138772 SEQ ID NO: 26 AAVH6 167 US20030138772 SEQ ID NO: 25AAVhE1.1 168 U.S. Pat. No. 9,233,131 SEQ ID NO: 44 AAVhEr1.14 169 U.S.Pat. No. 9,233,131 SEQ ID NO: 46 AAVhEr1.16 170 U.S. Pat. No. 9,233,131SEQ ID NO: 48 AAVhEr1.18 171 U.S. Pat. No. 9,233,131 SEQ ID NO: 49AAVhEr1.23 (AAVhEr2.29) 172 U.S. Pat. No. 9,233,131 SEQ ID NO: 53AAVhEr1.35 173 U.S. Pat. No. 9,233,131 SEQ ID NO: 50 AAVhEr1.36 174 U.S.Pat. No. 9,233,131 SEQ ID NO: 52 AAVhEr1.5 175 U.S. Pat. No. 9,233,131SEQ ID NO: 45 AAVhEr1.7 176 U.S. Pat. No. 9,233,131 SEQ ID NO: 51AAVhEr1.8 177 U.S. Pat. No. 9,233,131 SEQ ID NO: 47 AAVhEr2.16 178 U.S.Pat. No. 9,233,131 SEQ ID NO: 55 AAVhEr2.30 179 U.S. Pat. No. 9,233,131SEQ ID NO: 56 AAVhEr2.31 180 U.S. Pat. No. 9,233,131 SEQ ID NO: 58AAVhEr2.36 181 U.S. Pat. No. 9,233,131 SEQ ID NO: 57 AAVhEr2.4 182 U.S.Pat. No. 9,233,131 SEQ ID NO: 54 AAVhEr3.1 183 U.S. Pat. No. 9,233,131SEQ ID NO: 59 AAVhu.1 184 US20150315612 SEQ ID NO: 46 AAVhu.1 185US20150315612 SEQ ID NO: 144 AAVhu.10 (AAV16.8) 186 US20150315612 SEQ IDNO: 56 AAVhu.10 (AAV16.8) 187 US20150315612 SEQ ID NO: 156 AAVhu.11(AAV16.12) 188 US20150315612 SEQ ID NO: 57 AAVhu.11 (AAV16.12) 189US20150315612 SEQ ID NO: 153 AAVhu.12 190 US20150315612 SEQ ID NO: 59AAVhu.12 191 US20150315612 SEQ ID NO: 154 AAVhu.13 192 US20150159173 SEQID NO: 16, US20150315612 SEQ ID NO: 71 AAVhu.13 193 US20150159173 SEQ IDNO: 32, US20150315612 SEQ ID NO: 129 AAVhu.136.1 194 US20150315612 SEQID NO: 165 AAVhu.140.1 195 US20150315612 SEQ ID NO: 166 AAVhu.140.2 196US20150315612 SEQ ID NO: 167 AAVhu.145.6 197 US20150315612 SEQ ID No:178 AAVhu.15 198 US20150315612 SEQ ID NO: 147 AAVhu.15 (AAV33.4) 199US20150315612 SEQ ID NO: 50 AAVhu.156.1 200 US20150315612 SEQ ID No: 179AAVhu.16 201 US20150315612 SEQ ID NO: 148 AAVhu.16 (AAV33.8) 202US20150315612 SEQ ID NO: 51 AAVhu.17 203 US20150315612 SEQ ID NO: 83AAVhu.17 (AAV33.12) 204 US20150315612 SEQ ID NO: 4 AAVhu.172.1 205US20150315612 SEQ ID NO: 171 AAVhu.172.2 206 US20150315612 SEQ ID NO:172 AAVhu.173.4 207 US20150315612 SEQ ID NO: 173 AAVhu.173.8 208US20150315612 SEQ ID NO: 175 AAVhu.18 209 US20150315612 SEQ ID NO: 52AAVhu.18 210 US20150315612 SEQ ID NO: 149 AAVhu.19 211 US20150315612 SEQID NO: 62 AAVhu.19 212 US20150315612 SEQ ID NO: 133 AAVhu.2 213US20150315612 SEQ ID NO: 48 AAVhu.2 214 US20150315612 SEQ ID NO: 143AAVhu.20 215 US20150315612 SEQ ID NO: 63 AAVhu.20 216 US20150315612 SEQID NO: 134 AAVhu.21 217 US20150315612 SEQ ID NO: 65 AAVhu.21 218US20150315612 SEQ ID NO: 135 AAVhu.22 219 US20150315612 SEQ ID NO: 67AAVhu.22 220 US20150315612 SEQ ID NO: 138 AAVhu.23 221 US20150315612 SEQID NO: 60 AAVhu.23.2 222 US20150315612 SEQ ID NO: 137 AAVhu.24 223US20150315612 SEQ ID NO: 66 AAVhu.24 224 US20150315612 SEQ ID NO: 136AAVhu.25 225 US20150315612 SEQ ID NO: 49 AAVhu.25 226 US20150315612 SEQID NO: 146 AAVhu.26 227 US20150159173 SEQ ID NO: 17, US20150315612 SEQID NO: 61 AAVhu.26 228 US20150159173 SEQ ID NO: 33, US20150315612 SEQ IDNO: 139 AAVhu.27 229 US20150315612 SEQ ID NO: 64 AAVhu.27 230US20150315612 SEQ ID NO: 140 AAVhu.28 231 US20150315612 SEQ ID NO: 68AAVhu.28 232 US20150315612 SEQ ID NO: 130 AAVhu.29 233 US20150315612 SEQID NO: 69 AAVhu.29 234 US20150159173 SEQ ID NO: 42, US20150315612 SEQ IDNO: 132 AAVhu.29 235 US20150315612 SEQ ID NO: 225 AAVhu.29R 236US20150159173 AAVhu.3 237 US20150315612 SEQ ID NO: 44 AAVhu.3 238US20150315612 SEQ ID NO: 145 AAVhu.30 239 US20150315612 SEQ ID NO: 70AAVhu.30 240 US20150315612 SEQ ID NO: 131 AAVhu.31 241 US20150315612 SEQID NO: 1 AAVhu.31 242 US20150315612 SEQ ID NO: 121 AAVhu.32 243US20150315612 SEQ ID NO: 2 AAVhu.32 244 US20150315612 SEQ ID NO: 122AAVhu.33 245 US20150315612 SEQ ID NO: 75 AAVhu.33 246 US20150315612 SEQID NO: 124 AAVhu.34 247 US20150315612 SEQ ID NO: 72 AAVhu.34 248US20150315612 SEQ ID NO: 125 AAVhu.35 249 US20150315612 SEQ ID NO: 73AAVhu.35 250 US20150315612 SEQ ID NO: 164 AAVhu.36 251 US20150315612 SEQID NO: 74 AAVhu.36 252 US20150315612 SEQ ID NO: 126 AAVhu.37 253US20150159173 SEQ ID NO: 34, US20150315612 SEQ ID NO: 88 AAVhu.37(AAV106.1) 254 US20150315612 SEQ ID NO: 10, US20150159173 SEQ ID NO: 18AAVhu.38 255 US20150315612 SEQ ID NO: 161 AAVhu.39 256 US20150315612 SEQID NO: 102 AAVhu.39 (AAVLG-9) 257 US20150315612 SEQ ID NO: 24 AAVhu.4258 US20150315612 SEQ ID NO: 47 AAVhu.4 259 US20150315612 SEQ ID NO: 141AAVhu.40 260 US20150315612 SEQ ID NO: 87 AAVhu.40 (AAV114.3) 261US20150315612 SEQ ID No: 11 AAVhu.41 262 US20150315612 SEQ ID NO: 91AAVhu.41 (AAV127.2) 263 US20150315612 SEQ ID NO: 6 AAVhu.42 264US20150315612 SEQ ID NO: 85 AAVhu.42 (AAV127.5) 265 US20150315612 SEQ IDNO: 8 AAVhu.43 266 US20150315612 SEQ ID NO: 160 AAVhu.43 267US20150315612 SEQ ID NO: 236 AAVhu.43 (AAV128.1) 268 US20150315612 SEQID NO: 80 AAVhu.44 269 US20150159173 SEQ ID NO: 45, US20150315612 SEQ IDNO: 158 AAVhu.44 (AAV128.3) 270 US20150315612 SEQ ID NO: 81 AAVhu.44R1271 US20150159173 AAVhu.44R2 272 US20150159173 AAVhu.44R3 273US20150159173 AAVhu.45 274 US20150315612 SEQ ID NO: 76 AAVhu.45 275US20150315612 SEQ ID NO: 127 AAVhu.46 276 US20150315612 SEQ ID NO: 82AAVhu.46 277 US20150315612 SEQ ID NO: 159 AAVhu.46 278 US20150315612 SEQID NO: 224 AAVhu.47 279 US20150315612 SEQ ID NO: 77 AAVhu.47 280US20150315612 SEQ ID NO: 128 AAVhu.48 281 US20150159173 SEQ ID NO: 38AAVhu.48 282 US20150315612 SEQ ID NO: 157 AAVhu.48 (AAV130.4) 283US20150315612 SEQ ID NO: 78 AAVhu.48R1 284 US20150159173 AAVhu.48R2 285US20150159173 AAVhu.48R3 286 US20150159173 AAVhu.49 287 US20150315612SEQ ID NO: 209 AAVhu.49 288 US20150315612 SEQ ID NO: 189 AAVhu.5 289US20150315612 SEQ ID NO: 45 AAVhu.5 290 US20150315612 SEQ ID NO: 142AAVhu.51 291 US20150315612 SEQ ID NO: 208 AAVhu.51 292 US20150315612 SEQID NO: 190 AAVhu.52 293 US20150315612 SEQ ID NO: 210 AAVhu.52 294US20150315612 SEQ ID NO: 191 AAVhu.53 295 US20150159173 SEQ ID NO: 19AAVhu.53 296 US20150159173 SEQ ID NO: 35 AAVhu.53 (AAV145.1) 297US20150315612 SEQ ID NO: 176 AAVhu.54 298 US20150315612 SEQ ID NO: 188AAVhu.54 (AAV145.5) 299 US20150315612 SEQ ID No: 177 AAVhu.55 300US20150315612 SEQ ID NO: 187 AAVhu.56 301 US20150315612 SEQ ID NO: 205AAVhu.56 (AAV145.6) 302 US20150315612 SEQ ID NO: 168 AAVhu.56 (AAV145.6)303 US20150315612 SEQ ID NO: 192 AAVhu.57 304 US20150315612 SEQ ID NO:206 AAVhu.57 305 US20150315612 SEQ ID NO: 169 AAVhu.57 306 US20150315612SEQ ID NO: 193 AAVhu.58 307 US20150315612 SEQ ID NO: 207 AAVhu.58 308US20150315612 SEQ ID NO: 194 AAVhu.6 (AAV3.1) 309 US20150315612 SEQ IDNO: 5 AAVhu.6 (AAV3.1) 310 US20150315612 SEQ ID NO: 84 AAVhu.60 311US20150315612 SEQ ID NO: 184 AAVhu.60 (AAV161.10) 312 US20150315612 SEQID NO: 170 AAVhu.61 313 US20150315612 SEQ ID NO: 185 AAVhu.61 (AAV161.6)314 US20150315612 SEQ ID NO: 174 AAVhu.63 315 US20150315612 SEQ ID NO:204 AAVhu.63 316 US20150315612 SEQ ID NO: 195 AAVhu.64 317 US20150315612SEQ ID NO: 212 AAVhu.64 318 US20150315612 SEQ ID NO: 196 AAVhu.66 319US20150315612 SEQ ID NO: 197 AAVhu.67 320 US20150315612 SEQ ID NO: 215AAVhu.67 321 US20150315612 SEQ ID NO: 198 AAVhu.7 322 US20150315612 SEQID NO: 226 AAVhu.7 323 US20150315612 SEQ ID NO: 150 AAVhu.7 (AAV7.3) 324US20150315612 SEQ ID NO: 55 AAVhu.71 325 US20150315612 SEQ ID NO: 79AAVhu.8 326 US20150315612 SEQ ID NO: 53 AAVhu.8 327 US20150315612 SEQ IDNO: 12 AAVhu.8 328 US20150315612 SEQ ID NO: 151 AAVhu.9 (AAV3.1) 329US20150315612 SEQ ID NO: 58 AAVhu.9 (AAV3.1) 330 US20150315612 SEQ IDNO: 155 AAV-LK01 331 US20150376607 SEQ ID NO: 2 AAV-LK01 332US20150376607 SEQ ID NO: 29 AAV-LK02 333 US20150376607 SEQ ID NO: 3AAV-LK02 334 US20150376607 SEQ ID NO: 30 AAV-LK03 335 US20150376607 SEQID NO: 4 AAV-LK03 336 WO2015121501 SEQ ID NO: 12, US20150376607 SEQ IDNO: 31 AAV-LK04 337 US20150376607 SEQ ID NO: 5 AAV-LK04 338US20150376607 SEQ ID NO: 32 AAV-LK05 339 US20150376607 SEQ ID NO: 6AAV-LK05 340 US20150376607 SEQ ID NO: 33 AAV-LK06 341 US20150376607 SEQID NO: 7 AAV-LK06 342 US20150376607 SEQ ID NO: 34 AAV-LK07 343US20150376607 SEQ ID NO: 8 AAV-LK07 344 US20150376607 SEQ ID NO: 35AAV-LK08 345 US20150376607 SEQ ID NO: 9 AAV-LK08 346 US20150376607 SEQID NO: 36 AAV-LK09 347 US20150376607 SEQ ID NO: 10 AAV-LK09 348US20150376607 SEQ ID NO: 37 AAV-LK10 349 US20150376607 SEQ ID NO: 11AAV-LK10 350 US20150376607 SEQ ID NO: 38 AAV-LK11 351 US20150376607 SEQID NO: 12 AAV-LK11 352 US20150376607 SEQ ID NO: 39 AAV-LK12 353US20150376607 SEQ ID NO: 13 AAV-LK12 354 US20150376607 SEQ ID NO: 40AAV-LK13 355 US20150376607 SEQ ID NO: 14 AAV-LK13 356 US20150376607 SEQID NO: 41 AAV-LK14 357 US20150376607 SEQ ID NO: 15 AAV-LK14 358US20150376607 SEQ ID NO: 42 AAV-LK15 359 US20150376607 SEQ ID NO: 16AAV-LK15 360 US20150376607 SEQ ID NO: 43 AAV-LK16 361 US20150376607 SEQID NO: 17 AAV-LK16 362 US20150376607 SEQ ID NO: 44 AAV-LK17 363US20150376607 SEQ ID NO: 18 AAV-LK17 364 US20150376607 SEQ ID NO: 45AAV-LK18 365 US20150376607 SEQ ID NO: 19 AAV-LK18 366 US20150376607 SEQID NO: 46 AAV-LK19 367 US20150376607 SEQ ID NO: 20 AAV-LK19 368US20150376607 SEQ ID NO: 47 AAV-PAEC 369 US20150376607 SEQ ID NO: 1AAV-PAEC 370 US20150376607 SEQ ID NO: 48 AAV-PAEC11 371 US20150376607SEQ ID NO: 26 AAV-PAEC11 372 US20150376607 SEQ ID NO: 54 AAV-PAEC12 373US20150376607 SEQ ID NO: 27 AAV-PAEC12 374 US20150376607 SEQ ID NO: 51AAV-PAEC13 375 US20150376607 SEQ ID NO: 28 AAV-PAEC13 376 US20150376607SEQ ID NO: 49 AAV-PAEC2 377 US20150376607 SEQ ID NO: 21 AAV-PAEC2 378US20150376607 SEQ ID NO: 56 AAV-PAEC4 379 US20150376607 SEQ ID NO: 22AAV-PAEC4 380 US20150376607 SEQ ID NO: 55 AAV-PAEC6 381 US20150376607SEQ ID NO: 23 AAV-PAEC6 382 US20150376607 SEQ ID NO: 52 AAV-PAEC7 383US20150376607 SEQ ID NO: 24 AAV-PAEC7 384 US20150376607 SEQ ID NO: 53AAV-PAEC8 385 US20150376607 SEQ ID NO: 25 AAV-PAEC8 386 US20150376607SEQ ID NO: 50 AAVpi.1 387 US20150315612 SEQ ID NO: 28 AAVpi.1 388US20150315612 SEQ ID NO: 93 AAVpi.2 389 US20150315612 SEQ ID NO: 30AAVpi.2 390 US20150315612 SEQ ID NO: 95 AAVpi.3 391 US20150315612 SEQ IDNO: 29 AAVpi.3 392 US20150315612 SEQ ID NO: 94 AAVrh.10 393US20150159173 SEQ ID NO: 9 AAVrh.10 394 US20150159173 SEQ ID NO: 25AAV44.2 395 US20030138772 SEQ ID NO: 59 AAVrh.10 (AAV44.2) 396US20030138772 SEQ ID NO: 81 AAV42.1B 397 US20030138772 SEQ ID NO: 90AAVrh.12 (AAV42.1b) 398 US20030138772 SEQ ID NO: 30 AAVrh.13 399US20150159173 SEQ ID NO: 10 AAVrh.13 400 US20150159173 SEQ ID NO: 26AAVrh.13 401 US20150315612 SEQ ID NO: 228 AAVrh.l3R 402 US20150159173AAV42.3A 403 US20030138772 SEQ ID NO: 87 AAVrh.14 (AAV42.3a) 404US20030138772 SEQ ID NO: 32 AAV42.5A 405 US20030138772 SEQ ID NO: 89AAVrh.17 (AAV42.5a) 406 US20030138772 SEQ ID NO: 34 AAV42.5B 407US20030138772 SEQ ID NO: 91 AAVrh.18 (AAV42.5b) 408 US20030138772 SEQ IDNO: 29 AAV42.6B 409 US20030138772 SEQ ID NO: 112 AAVrh.19 (AAV42.6b) 410US20030138772 SEQ ID NO: 38 AAVrh.2 411 US20150159173 SEQ ID NO: 39AAVrh.2 412 US20150315612 SEQ ID NO: 231 AAVrh.20 413 US20150159173 SEQID NO: 1 AAV42.10 414 US20030138772 SEQ ID NO: 106 AAVrh.21 (AAV42.10)415 US20030138772 SEQ ID NO: 35 AAV42.11 416 US20030138772 SEQ ID NO:108 AAVrh.22 (AAV42.11) 417 US20030138772 SEQ ID NO: 37 AAV42.12 418US20030138772 SEQ ID NO: 113 AAVrh.23 (AAV42.12) 419 US20030138772 SEQID NO: 58 AAV42.13 420 US20030138772 SEQ ID NO: 86 AAVrh.24 (AAV42.13)421 US20030138772 SEQ ID NO: 31 AAV42.15 422 US20030138772 SEQ ID NO: 84AAVrh.25 (AAV42.15) 423 US20030138772 SEQ ID NO: 28 AAVrh.2R 424US20150159173 AAVrh.31 (AAV223.1) 425 US20030138772 SEQ ID NO: 48 AAVC1426 US20030138772 SEQ ID NO: 60 AAVrh.32 (AAVC1) 427 US20030138772 SEQID NO: 19 AAVrh.32/33 428 US20150159173 SEQ ID NO: 2 AAVrh.33 (AAVC3)429 US20030138772 SEQ ID NO: 20 AAVC5 430 US20030138772 SEQ ID NO: 62AAVrh.34 (AAVC5) 431 US20030138772 SEQ ID NO: 21 AAVF1 432 US20030138772SEQ ID NO: 109 AAVrh.35 (AAVF1) 433 US20030138772 SEQ ID NO: 22 AAVF3434 US20030138772 SEQ ID NO: 111 AAVrh.36 (AAVF3) 435 US20030138772 SEQID NO: 23 AAVrh.37 436 US20030138772 SEQ ID NO: 24 AAVrh.37 437US20150159173 SEQ ID NO: 40 AAVrh.37 438 US20150315612 SEQ ID NO: 229AAVrh.37R2 439 US20150159173 AAVrh.38 (AAVLG-4) 440 US20150315612 SEQ IDNO: 7 AAVrh.38 (AAVLG-4) 441 US20150315612 SEQ ID NO: 86 AAVrh.39 442US20150159173 SEQ ID NO: 20, US20150315612 SEQ ID NO: 13 AAVrh.39 443US20150159173 SEQ ID NO: 3, US20150159173 SEQ ID NO: 36, US20150315612SEQ ID NO: 89 AAVrh.40 444 US20150315612 SEQ ID NO: 92 AAVrh.40(AAVLG-10) 445 US20150315612 SEQ ID No: 14 AAVrh.43 (AAVN721-8) 446US20150315612 SEQ ID NO: 43, US20150159173 SEQ ID NO: 21 AAVrh.43(AAVN721-8) 447 US20150315612 SEQ ID NO: 163, US20150159173 SEQ ID NO:37 AAVrh.44 448 US20150315612 SEQ ID NO: 34 AAVrh.44 449 US20150315612SEQ ID NO: 111 AAVrh.45 450 US20150315612 SEQ ID NO: 41 AAVrh.45 451US20150315612 SEQ ID NO: 109 AAVrh.46 452 US20150159173 SEQ ID NO: 22,US20150315612 SEQ ID NO: 19 AAVrh.46 453 US20150159173 SEQ ID NO: 4,US20150315612 SEQ ID NO: 101 AAVrh.47 454 US20150315612 SEQ ID NO: 38AAVrh.47 455 US20150315612 SEQ ID NO: 118 AAVrh.48 456 US20150159173 SEQID NO: 44, US20150315612 SEQ ID NO: 115 AAVrh.48.1 457 US20150159173AAVrh.48.1.2 458 US20150159173 AAVrh.48.2 459 US20150159173 AAVrh.48(AAV1-7) 460 US20150315612 SEQ ID NO: 32 AAVrh.49 (AAV1-8) 461US20150315612 SEQ ID NO: 25 AAVrh.49 (AAV1-8) 462 US20150315612 SEQ IDNO: 103 AAVrh.50 (AAV2-4) 463 US20150315612 SEQ ID NO: 23 AAVrh.50(AAV2-4) 464 US20150315612 SEQ ID NO: 108 AAVrh.51 (AAV2-5) 465US20150315612 SEQ ID No: 22 AAVrh.51 (AAV2-5) 466 US20150315612 SEQ IDNO: 104 AAVrh.52 (AAV3-9) 467 US20150315612 SEQ ID NO: 18 AAVrh.52(AAV3-9) 468 US20150315612 SEQ ID NO: 96 AAVrh.53 469 US20150315612 SEQID NO: 97 AAVrh.53 (AAV3-11) 470 US20150315612 SEQ ID NO: 17 AAVrh.53(AAV3-11) 471 US20150315612 SEQ ID NO: 186 AAVrh.54 472 US20150315612SEQ ID NO: 40 AAVrh.54 473 US20150159173 SEQ ID NO: 49, US20150315612SEQ ID NO: 116 AVrh.55 474 US20150315612 SEQ ID NO: 37 AAVrh.55(AAV4-19) 475 US20150315612 SEQ ID NO: 117 AAVrh.56 476 US20150315612SEQ ID NO: 54 AAVrh.56 477 US20150315612 SEQ ID NO: 152 AAVrh.57 478US20150315612 SEQ ID NO: 26 AAVrh.57 479 US20150315612 SEQ ID NO: 105AAVrh.58 480 US20150315612 SEQ ID NO: 27 AAVrh.58 481 US20150159173 SEQID NO: 48, US20150315612 SEQ ID NO: 106 AAVrh.58 482 US20150315612 SEQID NO: 232 AAVrh.59 483 US20150315612 SEQ ID NO: 42 AAVrh.59 484US20150315612 SEQ ID NO: 110 AAVrh.60 485 US20150315612 SEQ ID NO: 31AAVrh.60 486 US20150315612 SEQ ID NO: 120 AAVrh.61 487 US20150315612 SEQID NO: 107 AAVrh.61 (AAV2-3) 488 US20150315612 SEQ ID NO: 21 AAVrh.62(AAV2-15) 489 US20150315612 SEQ ID No: 33 AAVrh.62 (AAV2-15) 490US20150315612 SEQ ID NO: 114 AAVrh.64 491 US20150315612 SEQ ID No: 15AAVrh.64 492 US20150159173 SEQ ID NO: 43, US20150315612 SEQ ID NO: 99AAVrh.64 493 US20150315612 SEQ ID NO: 233 AAVRh.64Rl 494 US20150159173AAVRh.64R2 495 US20150159173 AAVrh.65 496 US20150315612 SEQ ID NO: 35AAVrh.65 497 US20150315612 SEQ ID NO: 112 AAVrh.67 498 US20150315612 SEQID NO: 36 AAVrh.67 499 US20150315612 SEQ ID NO: 230 AAVrh.67 500US20150159173 SEQ ID NO: 47, US20150315612 SEQ ID NO: 113 AAVrh.68 501US20150315612 SEQ ID NO: 16 AAVrh.68 502 US20150315612 SEQ ID NO: 100AAVrh.69 503 US20150315612 SEQ ID NO: 39 AAVrh.69 504 US20150315612 SEQID NO: 119 AAVrh.70 505 US20150315612 SEQ ID NO: 20 AAVrh.70 506US20150315612 SEQ ID NO: 98 AAVrh.71 507 US20150315612 SEQ ID NO: 162AAVrh.72 508 US20150315612 SEQ ID NO: 9 AAVrh.73 509 US20150159173 SEQID NO: 5 AAVrh.74 510 US20150159173 SEQ ID NO: 6 AAVrh.8 511US20150159173 SEQ ID NO: 41 AAVrh.8 512 US20150315612 SEQ ID NO: 235AAVrh.8R 513 US20150159173, WO2015168666 SEQ ID NO: 9 AAVrh.8R A586Rmutant 514 WO2015168666 SEQ ID NO: 10 AAVrh.8R R533A mutant 515WO2015168666 SEQ ID NO: 11 BAAV (bovine AAV) 516 U.S. Pat. No. 9,193,769SEQ ID NO: 8 BAAV (bovine AAV) 517 U.S. Pat. No. 9,193,769 SEQ ID NO: 10BAAV (bovine AAV) 518 U.S. Pat. No. 9,193,769 SEQ ID NO: 4 BAAV (bovineAAV) 519 U.S. Pat. No. 9,193,769 SEQ ID NO: 2 BAAV (bovine AAV) 520 U.S.Pat. No. 9,193,769 SEQ ID NO: 6 BAAV (bovine AAV) 521 U.S. Pat. No.9,193,769 SEQ ID NO: 1 BAAV (bovine AAV) 522 U.S. Pat. No. 9,193,769 SEQID NO: 5 BAAV (bovine AAV) 523 U.S. Pat. No. 9,193,769 SEQ ID NO: 3 BAAV(bovine AAV) 524 U.S. Pat. No. 9,193,769 SEQ ID NO: 11 BAAV (bovine AAV)525 U.S. Pat. No. 7,427,396 SEQ ID NO: 5 BAAV (bovine AAV) 526 U.S. Pat.No. 7,427,396 SEQ ID NO: 6 BAAV (bovine AAV) 527 U.S. Pat. No. 9,193,769SEQ ID NO: 7 BAAV (bovine AAV) 528 U.S. Pat. No. 9,193,769 SEQ ID NO: 9BNP61 AAV 529 US20150238550 SEQ ID NO: 1 BNP61 AAV 530 US20150238550 SEQID NO: 2 BNP62 AAV 531 US20150238550 SEQ ID NO: 3 BNP63 AAV 532US20150238550 SEQ ID NO: 4 caprine AAV 533 U.S. Pat. No. 7,427,396 SEQID NO: 3 caprine AAV 534 U.S. Pat. No. 7,427,396 SEQ ID NO: 4 true typeAAV (ttAAV) 535 WO2015121501 SEQ ID NO: 2 AAAV (Avian AAV) 536 U.S. Pat.No. 7,427,396 SEQ ID NO: 12 AAAV (Avian AAV) 537 U.S. Pat. No. 9,238,800SEQ ID NO: 2 AAAV (Avian AAV) 538 U.S. Pat. No. 9,238,800 SEQ ID NO: 6AAAV (Avian AAV) 539 U.S. Pat. No. 9,238,800 SEQ ID NO: 4 AAAV (AvianAAV) 540 U.S. Pat. No. 9,238,800 SEQ ID NO: 8 AAAV (Avian AAV) 541 U.S.Pat. No. 9,238,800 SEQ ID NO: 14 AAAV (Avian AAV) 542 U.S. Pat. No.9,238,800 SEQ ID NO: 10 AAAV (Avian AAV) 543 U.S. Pat. No. 9,238,800 SEQID NO: 15 AAAV (Avian AAV) 544 U.S. Pat. No. 9,238,800 SEQ ID NO: 5 AAAV(Avian AAV) 545 U.S. Pat. No. 9,238,800 SEQ ID NO: 9 AAAV (Avian AAV)546 U.S. Pat. No. 9,238,800 SEQ ID NO: 3 AAAV (Avian AAV) 547 U.S. Pat.No. 9,238,800 SEQ ID NO: 7 AAAV (Avian AAV) 548 U.S. Pat. No. 9,238,800SEQ ID NO: 11 AAAV (Avian AAV) 549 U.S. Pat. No. 9,238,800 SEQ ID NO: 13AAAV (Avian AAV) 550 U.S. Pat. No. 9,238,800 SEQ ID NO: 1 AAV Shuffle100-1 551 US20160017295 SEQ ID NO: 23 AAV Shuffle 100-1 552US20160017295 SEQ ID NO: 11 AAV Shuffle 100-2 553 US20160017295 SEQ IDNO: 37 AAV Shuffle 100-2 554 US20160017295 SEQ ID NO: 29 AAV Shuffle100-3 555 US20160017295 SEQ ID NO: 24 AAV Shuffle 100-3 556US20160017295 SEQ ID NO: 12 AAV Shuffle 100-7 557 US20160017295 SEQ IDNO: 25 AAV Shuffle 100-7 558 US20160017295 SEQ ID NO: 13 AAV Shuffle10-2 559 US20160017295 SEQ ID NO: 34 AAV Shuffle 10-2 560 US20160017295SEQ ID NO: 26 AAV Shuffle 10-6 561 US20160017295 SEQ ID NO: 35 AAVShuffle 10-6 562 US20160017295 SEQ ID NO: 27 AAV Shuffle 10-8 563US20160017295 SEQ ID NO: 36 AAV Shuffle 10-8 564 US20160017295 SEQ IDNO: 28 AAV SM 100-10 565 US20160017295 SEQ ID NO: 41 AAV SM 100-10 566US20160017295 SEQ ID NO: 33 AAV SM 100-3 567 US20160017295 SEQ ID NO: 40AAV SM 100-3 568 US20160017295 SEQ ID NO: 32 AAV SM 10-1 569US20160017295 SEQ ID NO: 38 AAV SM 10-1 570 US20160017295 SEQ ID NO: 30AAV SM 10-2 571 US20160017295 SEQ ID NO: 10 AAV SM 10-2 572US20160017295 SEQ ID NO: 22 AAV SM 10-8 573 US20160017295 SEQ ID NO: 39AAV SM 10-8 574 US20160017295 SEQ ID NO: 31 AAVF1/HSC1 575 WO2016049230SEQ ID NO: 20 AAVF2/HSC2 576 WO2016049230 SEQ ID NO: 21 AAVF3/HSC3 577WO2016049230 SEQ ID NO: 22 AAVF4/HSC4 578 WO2016049230 SEQ ID NO: 23AAVF5/HSC5 579 WO2016049230 SEQ ID NO: 25 AAVF6/HSC6 580 WO2016049230SEQ ID NO: 24 AAVF7/HSC7 581 WO2016049230 SEQ ID NO: 27 AAVF8/HSC8 582WO2016049230 SEQ ID NO: 28 AAVF9/HSC9 583 WO2016049230 SEQ ID NO: 29AAVF11/HSC11 584 WO2016049230 SEQ ID NO: 26 AAVF12/HSC12 585WO2016049230 SEQ ID NO: 30 AAVF13/HSC13 586 WO2016049230 SEQ ID NO: 31AAVF14/HSC14 587 WO2016049230 SEQ ID NO: 32 AAVF15/HSC15 588WO2016049230 SEQ ID NO: 33 AAVF16/HSC16 589 WO2016049230 SEQ ID NO: 34AAVF17/HSC17 590 WO2016049230 SEQ ID NO: 35 AAVF1/HSC1 591 WO2016049230SEQ ID NO: 2 AAVF2/HSC2 592 WO2016049230 SEQ ID NO: 3 AAVF3/HSC3 593WO2016049230 SEQ ID NO: 5 AAVF4/HSC4 594 WO2016049230 SEQ ID NO: 6AAVF5/HSC5 595 WO2016049230 SEQ ID NO: 11 AAVF6/HSC6 596 WO2016049230SEQ ID NO: 7 AAVF7/HSC7 597 WO2016049230 SEQ ID NO: 8 AAVF8/HSC8 598WO2016049230 SEQ ID NO: 9 AAVF9/HSC9 599 WO2016049230 SEQ ID NO: 10AAVF11/HSC11 600 WO2016049230 SEQ ID NO: 4 AAVF12/HSC12 601 WO2016049230SEQ ID NO: 12 AAVF13/HSC13 602 WO2016049230 SEQ ID NO: 14 AAVF14/HSC14603 WO2016049230 SEQ ID NO: 15 AAVF15/HSC15 604 WO2016049230 SEQ ID NO:16 AAVF16/HSC16 605 WO2016049230 SEQ ID NO: 17 AAVF17/HSC17 606WO2016049230 SEQ ID NO: 13 AAV CBr-E1 607 U.S. Pat. No. 8,734,809 SEQ IDNO: 13 AAV CBr-E2 608 U.S. Pat. No. 8,734,809 SEQ ID NO: 14 AAV CBr-E3609 U.S. Pat. No. 8,734,809 SEQ ID NO: 15 AAV CBr-E4 610 U.S. Pat. No.8,734,809 SEQ ID NO: 16 AAV CBr-E5 611 U.S. Pat. No. 8,734,809 SEQ IDNO: 17 AAV CBr-e5 612 U.S. Pat. No. 8,734,809 SEQ ID NO: 18 AAV CBr-E6613 U.S. Pat. No. 8,734,809 SEQ ID NO: 19 AAV CBr-E7 614 U.S. Pat. No.8,734,809 SEQ ID NO: 20 AAV CBr-E8 615 U.S. Pat. No. 8,734,809 SEQ IDNO: 21 AAV CLv-D1 616 U.S. Pat. No. 8,734,809 SEQ ID NO: 22 AAV CLv-D2617 U.S. Pat. No. 8,734,809 SEQ ID NO: 23 AAV CLv-D3 618 U.S. Pat. No.8,734,809 SEQ ID NO: 24 AAV CLv-D4 619 U.S. Pat. No. 8,734,809 SEQ IDNO: 25 AAV CLv-D5 620 U.S. Pat. No. 8,734,809 SEQ ID NO: 26 AAV CLv-D6621 U.S. Pat. No. 8,734,809 SEQ ID NO: 27 AAV CLv-D7 622 U.S. Pat. No.8,734,809 SEQ ID NO: 28 AAV CLv-D8 623 U.S. Pat. No. 8,734,809 SEQ IDNO: 29 AAV CLv-E1 624 U.S. Pat. No. 8,734,809 SEQ ID NO: 13 AAV CLv-R1625 U.S. Pat. No. 8,734,809 SEQ ID NO: 30 AAV CLv-R2 626 U.S. Pat. No.8,734,809 SEQ ID NO: 31 AAV CLv-R3 627 U.S. Pat. No. 8,734,809 SEQ IDNO: 32 AAV CLv-R4 628 U.S. Pat. No. 8,734,809 SEQ ID NO: 33 AAV CLv-R5629 U.S. Pat. No. 8,734,809 SEQ ID NO: 34 AAV CLv-R6 630 U.S. Pat. No.8,734,809 SEQ ID NO: 35 AAV CLv-R7 631 U.S. Pat. No. 8,734,809 SEQ IDNO: 36 AAV CLv-R8 632 U.S. Pat. No. 8,734,809 SEQ ID NO: 37 AAV CLv-R9633 U.S. Pat. No. 8,734,809 SEQ ID NO: 38 AAV CLg-F1 634 U.S. Pat. No.8,734,809 SEQ ID NO: 39 AAV CLg-F2 635 U.S. Pat. No. 8,734,809 SEQ IDNO: 40 AAV CLg-F3 636 U.S. Pat. No. 8,734,809 SEQ ID NO: 41 AAV CLg-F4637 U.S. Pat. No. 8,734,809 SEQ ID NO: 42 AAV CLg-F5 638 U.S. Pat. No.8,734,809 SEQ ID NO: 43 AAV CLg-F6 639 U.S. Pat. No. 8,734,809 SEQ IDNO: 43 AAV CLg-F7 640 U.S. Pat. No. 8,734,809 SEQ ID NO: 44 AAV CLg-F8641 U.S. Pat. No. 8,734,809 SEQ ID NO: 43 AAV CSp-1 642 U.S. Pat. No.8,734,809 SEQ ID NO: 45 AAV CSp-10 643 U.S. Pat. No. 8,734,809 SEQ IDNO: 46 AAV CSp-11 644 U.S. Pat. No. 8,734,809 SEQ ID NO: 47 AAV CSp-2645 U.S. Pat. No. 8,734,809 SEQ ID NO: 48 AAV CSp-3 646 U.S. Pat. No.8,734,809 SEQ ID NO: 49 AAV CSp-4 647 U.S. Pat. No. 8,734,809 SEQ ID NO:50 AAV CSp-6 648 U.S. Pat. No. 8,734,809 SEQ ID NO: 51 AAV CSp-7 649U.S. Pat. No. 8,734,809 SEQ ID NO: 52 AAV CSp-8 650 U.S. Pat. No.8,734,809 SEQ ID NO: 53 AAV CSp-9 651 U.S. Pat. No. 8,734,809 SEQ ID NO:54 AAV CHt-2 652 U.S. Pat. No. 8,734,809 SEQ ID NO: 55 AAV CHt-3 653U.S. Pat. No. 8,734,809 SEQ ID NO: 56 AAV CKd-1 654 U.S. Pat. No.8,734,809 SEQ ID NO: 57 AAV CKd-10 655 U.S. Pat. No. 8,734,809 SEQ IDNO: 58 AAV CKd-2 656 U.S. Pat. No. 8,734,809 SEQ ID NO: 59 AAV CKd-3 657U.S. Pat. No. 8,734,809 SEQ ID NO: 60 AAV CKd-4 658 U.S. Pat. No.8,734,809 SEQ ID NO: 61 AAV CKd-6 659 U.S. Pat. No. 8,734,809 SEQ ID NO:62 AAV CKd-7 660 U.S. Pat. No. 8,734,809 SEQ ID NO: 63 AAV CKd-8 661U.S. Pat. No. 8,734,809 SEQ ID NO: 64 AAV CLv-1 662 U.S. Pat. No.8,734,809 SEQ ID NO: 65 AAV CLv-12 663 U.S. Pat. No. 8,734,809 SEQ IDNO: 66 AAV CLv-13 664 U.S. Pat. No. 8,734,809 SEQ ID NO: 67 AAV CLv-2665 U.S. Pat. No. 8,734,809 SEQ ID NO: 68 AAV CLv-3 666 U.S. Pat. No.8,734,809 SEQ ID NO: 69 AAV CLv-4 667 U.S. Pat. No. 8,734,809 SEQ ID NO:70 AAV CLv-6 668 U.S. Pat. No. 8,734,809 SEQ ID NO: 71 AAV CLv-8 669U.S. Pat. No. 8,734,809 SEQ ID NO: 72 AAV CKd-B1 670 U.S. Pat. No.8,734,809 SEQ ID NO: 73 AAV CKd-B2 671 U.S. Pat. No. 8,734,809 SEQ IDNO: 74 AAV CKd-B3 672 U.S. Pat. No. 8,734,809 SEQ ID NO: 75 AAV CKd-B4673 U.S. Pat. No. 8,734,809 SEQ ID NO: 76 AAV CKd-B5 674 U.S. Pat. No.8,734,809 SEQ ID NO: 77 AAV CKd-B6 675 U.S. Pat. No. 8,734,809 SEQ IDNO: 78 AAV CKd-B7 676 U.S. Pat. No. 8,734,809 SEQ ID NO: 79 AAV CKd-B8677 U.S. Pat. No. 8,734,809 SEQ ID NO: 80 AAV CKd-H1 678 U.S. Pat. No.8,734,809 SEQ ID NO: 81 AAV CKd-H2 679 U.S. Pat. No. 8,734,809 SEQ IDNO: 82 AAV CKd-H3 680 U.S. Pat. No. 8,734,809 SEQ ID NO: 83 AAV CKd-H4681 U.S. Pat. No. 8,734,809 SEQ ID NO: 84 AAV CKd-H5 682 U.S. Pat. No.8,734,809 SEQ ID NO: 85 AAV CKd-H6 683 U.S. Pat. No. 8,734,809 SEQ IDNO: 77 AAV CHt-1 684 U.S. Pat. No. 8,734,809 SEQ ID NO: 86 AAV CLv1-1685 U.S. Pat. No. 8,734,809 SEQ ID NO: 171 AAV CLv1-2 686 U.S. Pat. No.8,734,809 SEQ ID NO: 172 AAV CLv1-3 687 U.S. Pat. No. 8,734,809 SEQ IDNO: 173 AAV CLv1-4 688 U.S. Pat. No. 8,734,809 SEQ ID NO: 174 AAV Clv1-7689 U.S. Pat. No. 8,734,809 SEQ ID NO: 175 AAV Clv1-8 690 U.S. Pat. No.8,734,809 SEQ ID NO: 176 AAV Clv1-9 691 U.S. Pat. No. 8,734,809 SEQ IDNO: 177 AAV Clv1-10 692 U.S. Pat. No. 8,734,809 SEQ ID NO: 178AAV.VR-355 693 U.S. Pat. No. 8,734,809 SEQ ID NO: 181 AAV.hu.48R3 694U.S. Pat. No. 8,734,809 SEQ ID NO: 183 AAV CBr-E1 695 U.S. Pat. No.8,734,809 SEQ ID NO: 87 AAV CBr-E2 696 U.S. Pat. No. 8,734,809 SEQ IDNO: 88 AAV CBr-E3 697 U.S. Pat. No. 8,734,809 SEQ ID NO: 89 AAV CBr-E4698 U.S. Pat. No. 8,734,809 SEQ ID NO: 90 AAV CBr-E5 699 U.S. Pat. No.8,734,809 SEQ ID NO: 91 AAV CBr-e5 700 U.S. Pat. No. 8,734,809 SEQ IDNO: 92 AAV CBr-E6 701 U.S. Pat. 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No. 8,734,809 SEQ IDNO: 160 AAV CHt-P2 773 WO2016065001 SEQ ID NO: 1 AAV CHt-P5 774WO2016065001 SEQ ID NO: 2 AAV CHt-P9 775 WO2016065001 SEQ ID NO: 3 AAVCBr-7.1 776 WO2016065001 SEQ ID NO: 4 AAV CBr-7.2 777 WO2016065001 SEQID NO: 5 AAV CBr-7.3 778 WO2016065001 SEQ ID NO: 6 AAV CBr-7.4 779WO2016065001 SEQ ID NO: 7 AAV CBr-7.5 780 WO2016065001 SEQ ID NO: 8 AAVCBr-7.7 781 WO2016065001 SEQ ID NO: 9 AAV CBr-7.8 782 WO2016065001 SEQID NO: 10 AAV CBr-7.10 783 WO2016065001 SEQ ID NO: 11 AAV CKd-N3 784WO2016065001 SEQ ID NO: 12 AAV CKd-N4 785 WO2016065001 SEQ ID NO: 13 AAVCKd-N9 786 WO2016065001 SEQ ID NO: 14 AAV CLv-L4 787 WO2016065001 SEQ IDNO: 15 AAV CLv-L5 788 WO2016065001 SEQ ID NO: 16 AAV CLv-L6 789WO2016065001 SEQ ID NO: 17 AAV CLv-K1 790 WO2016065001 SEQ ID NO: 18 AAVCLv-K3 791 WO2016065001 SEQ ID NO: 19 AAV CLv-K6 792 WO2016065001 SEQ IDNO: 20 AAV CLv-M1 793 WO2016065001 SEQ ID NO: 21 AAV CLv-M11 794WO2016065001 SEQ ID NO: 22 AAV CLv-M2 795 WO2016065001 SEQ ID NO: 23 AAVCLv-M5 796 WO2016065001 SEQ ID NO: 24 AAV CLv-M6 797 WO2016065001 SEQ IDNO: 25 AAV CLv-M7 798 WO2016065001 SEQ ID NO: 26 AAV CLv-M8 799WO2016065001 SEQ ID NO: 27 AAV CLv-M9 800 WO2016065001 SEQ ID NO: 28 AAVCHt-P1 801 WO2016065001 SEQ ID NO: 29 AAV CHt-P6 802 WO2016065001 SEQ IDNO: 30 AAV CHt-P8 803 WO2016065001 SEQ ID NO: 31 AAV CHt-6.1 804WO2016065001 SEQ ID NO: 32 AAV CHt-6.10 805 WO2016065001 SEQ ID NO: 33AAV CHt-6.5 806 WO2016065001 SEQ ID NO: 34 AAV CHt-6.6 807 WO2016065001SEQ ID NO: 35 AAV CHt-6.7 808 WO2016065001 SEQ ID NO: 36 AAV CHt-6.8 809WO2016065001 SEQ ID NO: 37 AAV CSp-8.10 810 WO2016065001 SEQ ID NO: 38AAV CSp-8.2 811 WO2016065001 SEQ ID NO: 39 AAV CSp-8.4 812 WO2016065001SEQ ID NO: 40 AAV CSp-8.5 813 WO2016065001 SEQ ID NO: 41 AAV CSp-8.6 814WO2016065001 SEQ ID NO: 42 AAV CSp-8.7 815 WO2016065001 SEQ ID NO: 43AAV CSp-8.8 816 WO2016065001 SEQ ID NO: 44 AAV CSp-8.9 817 WO2016065001SEQ ID NO: 45 AAV CBr-B7.3 818 WO2016065001 SEQ ID NO: 46 AAV CBr-B7.4819 WO2016065001 SEQ ID NO: 47 AAV3B 820 WO2016065001 SEQ ID NO: 48 AAV4821 WO2016065001 SEQ ID NO: 49 AAV5 822 WO2016065001 SEQ ID NO: 50 AAVCHt-P2 823 WO2016065001 SEQ ID NO: 51 AAV CHt-P5 824 WO2016065001 SEQ IDNO: 52 AAV CHt-P9 825 WO2016065001 SEQ ID NO: 53 AAV CBr-7.1 826WO2016065001 SEQ ID NO: 54 AAV CBr-7.2 827 WO2016065001 SEQ ID NO: 55AAV CBr-7.3 828 WO2016065001 SEQ ID NO: 56 AAV CBr-7.4 829 WO2016065001SEQ ID NO: 57 AAV CBr-7.5 830 WO2016065001 SEQ ID NO: 58 AAV CBr-7.7 831WO2016065001 SEQ ID NO: 59 AAV CBr-7.8 832 WO2016065001 SEQ ID NO: 60AAV CBr-7.10 833 WO2016065001 SEQ ID NO: 61 AAV CKd-N3 834 WO2016065001SEQ ID NO: 62 AAV CKd-N4 835 WO2016065001 SEQ ID NO: 63 AAV CKd-N9 836WO2016065001 SEQ ID NO: 64 AAV CLv-L4 837 WO2016065001 SEQ ID NO: 65 AAVCLv-L5 838 WO2016065001 SEQ ID NO: 66 AAV CLv-L6 839 WO2016065001 SEQ IDNO: 67 AAV CLv-K1 840 WO2016065001 SEQ ID NO: 68 AAV CLv-K3 841WO2016065001 SEQ ID NO: 69 AAV CLv-K6 842 WO2016065001 SEQ ID NO: 70 AAVCLv-M1 843 WO2016065001 SEQ ID NO: 71 AAV CLv-M1 844 WO2016065001 SEQ IDNO: 72 AAV CLv-M2 845 WO2016065001 SEQ ID NO: 73 AAV CLv-M5 846WO2016065001 SEQ ID NO: 74 AAV CLv-M6 847 WO2016065001 SEQ ID NO: 75 AAVCLv-M7 848 WO2016065001 SEQ ID NO: 76 AAV CLv-M8 849 WO2016065001 SEQ IDNO: 77 AAV CLv-M9 850 WO2016065001 SEQ ID NO: 78 AAV CHt-P1 851WO2016065001 SEQ ID NO: 79 AAV CHt-P6 852 WO2016065001 SEQ ID NO: 80 AAVCHt-P8 853 WO2016065001 SEQ ID NO: 81 AAV CHt-6.1 854 WO2016065001 SEQID NO: 82 AAV CHt-6.10 855 WO2016065001 SEQ ID NO: 83 AAV CHt-6.5 856WO2016065001 SEQ ID NO: 84 AAV CHt-6.6 857 WO2016065001 SEQ ID NO: 85AAV CHt-6.7 858 WO2016065001 SEQ ID NO: 86 AAV CHt-6.8 859 WO2016065001SEQ ID NO: 87 AAV CSp-8.10 860 WO2016065001 SEQ ID NO: 88 AAV CSp-8.2861 WO2016065001 SEQ ID NO: 89 AAV CSp-8.4 862 WO2016065001 SEQ ID NO:90 AAV CSp-8.5 863 WO2016065001 SEQ ID NO: 91 AAV CSp-8.6 864WO2016065001 SEQ ID NO: 92 AAV CSp-8.7 865 WO2016065001 SEQ ID NO: 93AAV CSp-8.8 866 WO2016065001 SEQ ID NO: 94 AAV CSp-8.9 867 WO2016065001SEQ ID NO: 95 AAV CBr-B7.3 868 WO2016065001 SEQ ID NO: 96 AAV CBr-B7.4869 WO2016065001 SEQ ID NO: 97 AAV3B 870 WO2016065001 SEQ ID NO: 98 AAV4871 WO2016065001 SEQ ID NO: 99 AAV5 872 WO2016065001 SEQ ID NO: 100PHP.N/PHP.B-DGT 873 WO2017100671 SEQ ID NO: 46 PHP.S/G2A12 874WO2017100671 SEQ ID NO: 47 AAV9/hu.14 K449R 875 WO2017100671 SEQ ID NO:45 GPV 992 U.S. Pat. No. 9,624,274B2 SEQ ID NO: 192 B19 993 U.S. Pat.No. 9,624,274B2 SEQ ID NO: 193 MVM 994 U.S. Pat. No. 9,624,274B2 SEQ IDNO: 194 FPV 995 U.S. Pat. No. 9,624,274B2 SEQ ID NO: 195 CPV 996 U.S.Pat. No. 9,624,274B2 SEQ ID NO: 196 AAV6 997 U.S. Pat. No. 9,546,112B2SEQ ID NO: 5 AAV6 998 U.S. Pat. No. 9,457,103B2 SEQ ID NO: 1 AAV2 999U.S. Pat. No. 9,457,103B2 SEQ ID NO: 2 ShH10 1000 U.S. Pat. No.9,457,103B2 SEQ ID NO: 3 ShH13 1001 U.S. Pat. No. 9,457,103B2 SEQ ID NO:4 ShH10 1002 U.S. Pat. No. 9,457,103B2 SEQ ID NO: 5 ShH10 1003 U.S. Pat.No. 9,457,103B2 SEQ ID NO: 6 ShH10 1004 U.S. Pat. No. 9,457,103B2 SEQ IDNO: 7 ShH10 1005 U.S. Pat. No. 9,457,103B2 SEQ ID NO: 8 ShH10 1006 U.S.Pat. No. 9,457,103B2 SEQ ID NO: 9 rh74 1007 U.S. Pat. No. 9,434,928B2SEQ ID NO: 1, US2015023924A1 SEQ ID NO: 2 rh74 1008 U.S. Pat. No.9,434,928B2 SEQ ID NO: 2, US2015023924A1 SEQ ID NO: 1 AAV8 1009 U.S.Pat. No. 9,434,928B2 SEQ ID NO: 4 rh74 1010 U.S. Pat. No. 9,434,928B2SEQ ID NO: 5 rh74 (RHM4-1) 1011 US2015023924A1 SEQ ID NO: 5,US20160375110A1 SEQ ID NO: 4 rh74 (RHM15-1) 1012 US2015023924A1 SEQ IDNO: 6, US20160375110A1 SEQ ID NO: 5 rh74 (RHM15-2) 1013 US2015023924A1SEQ ID NO: 7, US20160375110A1 SEQ ID NO: 6 rh74 (RHM15-3/RHM15-5) 1014US2015023924A1 SEQ ID NO: 8, US20160375110A1 SEQ ID NO: 7 rh74 (RHM15-4)1015 US2015023924A1 SEQ ID NO: 9, US20160375110A1 SEQ ID NO: 8 rh74(RHM15-6) 1016 US2015023924A1 SEQ ID NO: 10, US20160375110A1 SEQ ID NO:9 rh74 (RHM4-1) 1017 US2015023924A1 SEQ ID NO: 11 rh74 (RHM15-1) 1018US2015023924A1 SEQ ID NO: 12 rh74 (RHM15-2) 1019 US2015023924A1 SEQ IDNO: 13 rh74 (RHM15-3/RHM15-5) 1020 US2015023924A1 SEQ ID NO: 14 rh74(RHM15-4) 1021 US2015023924A1 SEQ ID NO: 15 rh74 (RHM15-6) 1022US2015023924A1 SEQ ID NO: 16 AAV2 (comprising lung 1023 US20160175389A1SEQ ID NO: 9 specific polypeptide) AAV2 (comprising lung 1024US20160175389A1 SEQ ID NO: 10 specific polypeptide) Anc80 1025US20170051257A1 SEQ ID NO: 1 Anc80 1026 US20170051257A1 SEQ ID NO: 2Anc81 1027 US20170051257A1 SEQ ID NO: 3 Anc80 1028 US20170051257A1 SEQID NO: 4 Anc82 1029 US20170051257A1 SEQ ID NO: 5 Anc82 1030US20170051257A1 SEQ ID NO: 6 Anc83 1031 US20170051257A1 SEQ ID NO: 7Anc83 1032 US20170051257A1 SEQ ID NO: 8 Anc84 1033 US20170051257A1 SEQID NO: 9 Anc84 1034 US20170051257A1 SEQ ID NO: 10 Anc94 1035US20170051257A1 SEQ ID NO: 11 Anc94 1036 US20170051257A1 SEQ ID NO: 12Anc113 1037 US20170051257A1 SEQ ID NO: 13 Anc113 1038 US20170051257A1SEQ ID NO: 14 Anc126 1039 US20170051257A1 SEQ ID NO: 15 Anc126 1040US20170051257A1 SEQ ID NO: 16 Anc127 1041 US20170051257A1 SEQ ID NO: 17Anc127 1042 US20170051257A1 SEQ ID NO: 18 Anc80L27 1043 US20170051257A1SEQ ID NO: 19 Anc80L59 1044 US20170051257A1 SEQ ID NO: 20 Anc80L60 1045US20170051257A1 SEQ ID NO: 21 Anc80L62 1046 US20170051257A1 SEQ ID NO:22 Anc80L65 1047 US20170051257A1 SEQ ID NO: 23 Anc80L33 1048US20170051257A1 SEQ ID NO: 24 Anc80L36 1049 US20170051257A1 SEQ ID NO:25 Anc80L44 1050 US20170051257A1 SEQ ID NO: 26 Anc80L1 1051US20170051257A1 SEQ ID NO: 35 Anc80L1 1052 US20170051257A1 SEQ ID NO: 36AAV-X1 1053 U.S. Pat. No. 8,283,151B2 SEQ ID NO: 11 AAV-X1b 1054 U.S.Pat. No. 8,283,151B2 SEQ ID NO: 12 AAV-X5 1055 U.S. Pat. No. 8,283,151B2SEQ ID NO: 13 AAV-X19 1056 U.S. Pat. No. 8,283,151B2 SEQ ID NO: 14AAV-X21 1057 U.S. Pat. No. 8,283,151B2 SEQ ID NO: 15 AAV-X22 1058 U.S.Pat. No. 8,283,151B2 SEQ ID NO: 16 AAV-X23 1059 U.S. Pat. No.8,283,151B2 SEQ ID NO: 17 AAV-X24 1060 U.S. Pat. No. 8,283,151B2 SEQ IDNO: 18 AAV-X25 1061 U.S. Pat. No. 8,283,151B2 SEQ ID NO: 19 AAV-X26 1062U.S. Pat. No. 8,283,151B2 SEQ ID NO: 20 AAV-X1 1063 U.S. Pat. No.8,283,151B2 SEQ ID NO: 21 AAV-X1b 1064 U.S. Pat. No. 8,283,151B2 SEQ IDNO: 22 AAV-X5 1065 U.S. Pat. No. 8,283,151B2 SEQ ID NO: 23 AAV-X19 1066U.S. Pat. No. 8,283,151B2 SEQ ID NO: 24 AAV-X21 1067 U.S. Pat. No.8,283,151B2 SEQ ID NO: 25 AAV-X22 1068 U.S. Pat. No. 8,283,151B2 SEQ IDNO: 26 AAV-X23 1069 U.S. Pat. No. 8,283,151B2 SEQ ID NO: 27 AAV-X24 1070U.S. Pat. No. 8,283,151B2 SEQ ID NO: 28 AAV-X25 1071 U.S. Pat. No.8,283,151B2 SEQ ID NO: 29 AAV-X26 1072 U.S. Pat. No. 8,283,151B2 SEQ IDNO: 30 AAVrh8 1073 WO2016054554A1 SEQ ID NO: 8 AAVrh8VP2FC5 1074WO2016054554A1 SEQ ID NO: 9 AAVrh8VP2FC44 1075 WO2016054554A1 SEQ ID NO:10 AAVrh8VP2ApoB100 1076 WO2016054554A1 SEQ ID NO: 11 AAVrh8VP2RVG 1077WO2016054554A1 SEQ ID NO: 12 AAVrh8VP2Angiopep-2 1078 WO2016054554A1 SEQID NO: 13 VP2 AAV9.47VP1.3 1079 WO2016054554A1 SEQ ID NO: 14AAV9.47VP2ICAMg3 1080 WO2016054554A1 SEQ ID NO: 15 AAV9.47VP2RVG 1081WO2016054554A1 SEQ ID NO: 16 AAV9.47VP2Angiopep-2 1082 WO2016054554A1SEQ ID NO: 17 AAV9.47VP2A-string 1083 WO2016054554A1 SEQ ID NO: 18AAVrh8VP2FC5 VP2 1084 WO2016054554A1 SEQ ID NO: 19 AAVrh8VP2FC44 VP21085 WO2016054554A1 SEQ ID NO: 20 AAVrh8VP2ApoB100 VP2 1086WO2016054554A1 SEQ ID NO: 21 AAVrh8VP2RVG VP2 1087 WO2016054554A1 SEQ IDNO: 22 AAVrh8VP2Angiopep-2 1088 WO2016054554A1 SEQ ID NO: 23 VP2AAV9.47VP2ICAMg3 VP2 1089 WO2016054554A1 SEQ ID NO: 24 AAV9.47VP2RVG VP21090 WO2016054554A1 SEQ ID NO: 25 AAV9.47VP2Angiopep-2 1091WO2016054554A1 SEQ ID NO: 26 VP2 AAV9.47VP2A-string VP2 1092WO2016054554A1 SEQ ID NO: 27 rAAV-B1 1093 WO2016054557A1 SEQ ID NO: 1rAAV-B2 1094 WO2016054557A1 SEQ ID NO: 2 rAAV-B3 1095 WO2016054557A1 SEQID NO: 3 rAAV-B4 1096 WO2016054557A1 SEQ ID NO: 4 rAAV-B1 1097WO2016054557A1 SEQ ID NO: 5 rAAV-B2 1098 WO2016054557A1 SEQ ID NO: 6rAAV-B3 1099 WO2016054557A1 SEQ ID NO: 7 rAAV-B4 1100 WO2016054557A1 SEQID NO: 8 rAAV-L1 1101 WO2016054557A1 SEQ ID NO: 9 rAAV-L2 1102WO2016054557A1 SEQ ID NO: 10 rAAV-L3 1103 WO2016054557A1 SEQ ID NO: 11rAAV-L4 1104 WO2016054557A1 SEQ ID NO: 12 rAAV-L1 1105 WO2016054557A1SEQ ID NO: 13 rAAV-L2 1106 WO2016054557A1 SEQ ID NO: 14 rAAV-L3 1107WO2016054557A1 SEQ ID NO: 15 rAAV-L4 1108 WO2016054557A1 SEQ ID NO: 16AAV9 1109 WO2016073739A1 SEQ ID NO: 3 rAAV 1110 WO2016081811A1 SEQ IDNO: 1 rAAV 1111 WO2016081811A1 SEQ ID NO: 2 rAAV 1112 WO2016081811A1 SEQID NO: 3 rAAV 1113 WO2016081811A1 SEQ ID NO: 4 rAAV 1114 WO2016081811A1SEQ ID NO: 5 rAAV 1115 WO2016081811A1 SEQ ID NO: 6 rAAV 1116WO2016081811A1 SEQ ID NO: 7 rAAV 1117 WO2016081811A1 SEQ ID NO: 8 rAAV1118 WO2016081811A1 SEQ ID NO: 9 rAAV 1119 WO2016081811A1 SEQ ID NO: 10rAAV 1120 WO2016081811A1 SEQ ID NO: 11 rAAV 1121 WO2016081811A1 SEQ IDNO: 12 rAAV 1122 WO2016081811A1 SEQ ID NO: 13 rAAV 1123 WO2016081811A1SEQ ID NO: 14 rAAV 1124 WO2016081811A1 SEQ ID NO: 15 rAAV 1125WO2016081811A1 SEQ ID NO: 16 rAAV 1126 WO2016081811A1 SEQ ID NO: 17 rAAV1127 WO2016081811A1 SEQ ID NO: 18 rAAV 1128 WO2016081811A1 SEQ ID NO: 19rAAV 1129 WO2016081811A1 SEQ ID NO: 20 rAAV 1130 WO2016081811A1 SEQ IDNO: 21 rAAV 1131 WO2016081811A1 SEQ ID NO: 22 rAAV 1132 WO2016081811A1SEQ ID NO: 23 rAAV 1133 WO2016081811A1 SEQ ID NO: 24 rAAV 1134WO2016081811A1 SEQ ID NO: 25 rAAV 1135 WO2016081811A1 SEQ ID NO: 26 rAAV1136 WO2016081811A1 SEQ ID NO: 27 rAAV 1137 WO2016081811A1 SEQ ID NO: 28rAAV 1138 WO2016081811A1 SEQ ID NO: 29 rAAV 1139 WO2016081811A1 SEQ IDNO: 30 rAAV 1140 WO2016081811A1 SEQ ID NO: 31 rAAV 1141 WO2016081811A1SEQ ID NO: 32 rAAV 1142 WO2016081811A1 SEQ ID NO: 33 rAAV 1143WO2016081811A1 SEQ ID NO: 34 rAAV 1144 WO2016081811A1 SEQ ID NO: 35 rAAV1145 WO2016081811A1 SEQ ID NO: 36 rAAV 1146 WO2016081811A1 SEQ ID NO: 37rAAV 1147 WO2016081811A1 SEQ ID NO: 38 rAAV 1148 WO2016081811A1 SEQ IDNO: 39 rAAV 1149 WO2016081811A1 SEQ ID NO: 40 rAAV 1150 WO2016081811A1SEQ ID NO: 41 rAAV 1151 WO2016081811A1 SEQ ID NO: 42 rAAV 1152WO2016081811A1 SEQ ID NO: 43 rAAV 1153 WO2016081811A1 SEQ ID NO: 44 rAAV1154 WO2016081811A1 SEQ ID NO: 45 rAAV 1155 WO2016081811A1 SEQ ID NO: 46rAAV 1156 WO2016081811A1 SEQ ID NO: 47 rAAV 1157 WO2016081811A1 SEQ IDNO: 48 rAAV 1158 WO2016081811A1 SEQ ID NO: 49 rAAV 1159 WO2016081811A1SEQ ID NO: 50 rAAV 1160 WO2016081811A1 SEQ ID NO: 51 rAAV 1161WO2016081811A1 SEQ ID NO: 52 rAAV 1162 WO2016081811A1 SEQ ID NO: 53 rAAV1163 WO2016081811A1 SEQ ID NO: 54 rAAV 1164 WO2016081811A1 SEQ ID NO: 55rAAV 1165 WO2016081811A1 SEQ ID NO: 56 rAAV 1166 WO2016081811A1 SEQ IDNO: 57 rAAV 1167 WO2016081811A1 SEQ ID NO: 58 rAAV 1168 WO2016081811A1SEQ ID NO: 59 rAAV 1169 WO2016081811A1 SEQ ID NO: 60 rAAV 1170WO2016081811A1 SEQ ID NO: 61 rAAV 1171 WO2016081811A1 SEQ ID NO: 62 rAAV1172 WO2016081811A1 SEQ ID NO: 63 rAAV 1173 WO2016081811A1 SEQ ID NO: 64rAAV 1174 WO2016081811A1 SEQ ID NO: 65 rAAV 1175 WO2016081811A1 SEQ IDNO: 66 rAAV 1176 WO2016081811A1 SEQ ID NO: 67 rAAV 1177 WO2016081811A1SEQ ID NO: 68 rAAV 1178 WO2016081811A1 SEQ ID NO: 69 rAAV 1179WO2016081811A1 SEQ ID NO: 70 rAAV 1180 WO2016081811A1 SEQ ID NO: 71 rAAV1181 WO2016081811A1 SEQ ID NO: 72 rAAV 1182 WO2016081811A1 SEQ ID NO: 73rAAV 1183 WO2016081811A1 SEQ ID NO: 74 rAAV 1184 WO2016081811A1 SEQ IDNO: 75 rAAV 1185 WO2016081811A1 SEQ ID NO: 76 rAAV 1186 WO2016081811A1SEQ ID NO: 77 rAAV 1187 WO2016081811A1 SEQ ID NO: 78 rAAV 1188WO2016081811A1 SEQ ID NO: 79 rAAV 1189 WO2016081811A1 SEQ ID NO: 80 rAAV1190 WO2016081811A1 SEQ ID NO: 81 rAAV 1191 WO2016081811A1 SEQ ID NO: 82rAAV 1192 WO2016081811A1 SEQ ID NO: 83 rAAV 1193 WO2016081811A1 SEQ IDNO: 84 rAAV 1194 WO2016081811A1 SEQ ID NO: 85 rAAV 1195 WO2016081811A1SEQ ID NO: 86 rAAV 1196 WO2016081811A1 SEQ ID NO: 87 rAAV 1197WO2016081811A1 SEQ ID NO: 88 rAAV 1198 WO2016081811A1 SEQ ID NO: 89 rAAV1199 WO2016081811A1 SEQ ID NO: 90 rAAV 1200 WO2016081811A1 SEQ ID NO: 91rAAV 1201 WO2016081811A1 SEQ ID NO: 92 rAAV 1202 WO2016081811A1 SEQ IDNO: 93 rAAV 1203 WO2016081811A1 SEQ ID NO: 94 rAAV 1204 WO2016081811A1SEQ ID NO: 95 rAAV 1205 WO2016081811A1 SEQ ID NO: 96 rAAV 1206WO2016081811A1 SEQ ID NO: 97 rAAV 1207 WO2016081811A1 SEQ ID NO: 98 rAAV1208 WO2016081811A1 SEQ ID NO: 99 rAAV 1209 WO2016081811A1 SEQ ID NO:100 rAAV 1210 WO2016081811A1 SEQ ID NO: 101 rAAV 1211 WO2016081811A1 SEQID NO: 102 rAAV 1212 WO2016081811A1 SEQ ID NO: 103 rAAV 1213WO2016081811A1 SEQ ID NO: 104 rAAV 1214 WO2016081811A1 SEQ ID NO: 105rAAV 1215 WO2016081811A1 SEQ ID NO: 106 rAAV 1216 WO2016081811A1 SEQ IDNO: 107 rAAV 1217 WO2016081811A1 SEQ ID NO: 108 rAAV 1218 WO2016081811A1SEQ ID NO: 109 rAAV 1219 WO2016081811A1 SEQ ID NO: 110 rAAV 1220WO2016081811A1 SEQ ID NO: 111 rAAV 1221 WO2016081811A1 SEQ ID NO: 112rAAV 1222 WO2016081811A1 SEQ ID NO: 113 rAAV 1223 WO2016081811A1 SEQ IDNO: 114 rAAV 1224 WO2016081811A1 SEQ ID NO: 115 rAAV 1225 WO2016081811A1SEQ ID NO: 116 rAAV 1226 WO2016081811A1 SEQ ID NO: 117 rAAV 1227WO2016081811A1 SEQ ID NO: 118 rAAV 1228 WO2016081811A1 SEQ ID NO: 119rAAV 1229 WO2016081811A1 SEQ ID NO: 120 rAAV 1230 WO2016081811A1 SEQ IDNO: 121 rAAV 1231 WO2016081811A1 SEQ ID NO: 122 rAAV 1232 WO2016081811A1SEQ ID NO: 123 rAAV 1233 WO2016081811A1 SEQ ID NO: 124 rAAV 1234WO2016081811A1 SEQ ID NO: 125 rAAV 1235 WO2016081811A1 SEQ ID NO: 126rAAV 1236 WO2016081811A1 SEQ ID NO: 127 rAAV 1237 WO2016081811A1 SEQ IDNO: 128 AAV8 E532K 1238 WO2016081811A1 SEQ ID NO: 133 AAV8 E532K 1239WO2016081811A1 SEQ ID NO: 134 rAAV4 1240 WO2016115382A1 SEQ ID NO: 2rAAV4 1241 WO2016115382A1 SEQ ID NO: 3 rAAV4 1242 WO2016115382A1 SEQ IDNO: 4 rAAV4 1243 WO2016115382A1 SEQ ID NO: 5 rAAV4 1244 WO2016115382A1SEQ ID NO: 6 rAAV4 1245 WO2016115382A1 SEQ ID NO: 7 rAAV4 1246WO2016115382A1 SEQ ID NO: 8 rAAV4 1247 WO2016115382A1 SEQ ID NO: 9 rAAV41248 WO2016115382A1 SEQ ID NO: 10 rAAV4 1249 WO2016115382A1 SEQ ID NO:11 rAAV4 1250 WO2016115382A1 SEQ ID NO: 12 rAAV4 1251 WO2016115382A1 SEQID NO: 13 rAAV4 1252 WO2016115382A1 SEQ ID NO: 14 rAAV4 1253WO2016115382A1 SEQ ID NO: 15 rAAV4 1254 WO2016115382A1 SEQ ID NO: 16rAAV4 1255 WO2016115382A1 SEQ ID NO: 17 rAAV4 1256 WO2016115382A1 SEQ IDNO: 18 rAAV4 1257 WO2016115382A1 SEQ ID NO: 19 rAAV4 1258 WO2016115382A1SEQ ID NO: 20 rAAV4 1259 WO2016115382A1 SEQ ID NO: 21 AAV11 1260WO2016115382A1 SEQ ID NO: 22 AAV12 1261 WO2016115382A1 SEQ ID NO: 23rh32 1262 WO2016115382A1 SEQ ID NO: 25 rh33 1263 WO2016115382A1 SEQ IDNO: 26 rh34 1264 WO2016115382A1 SEQ ID NO: 27 rAAV4 1265 WO2016115382A1SEQ ID NO: 28 rAAV4 1266 WO2016115382A1 SEQ ID NO: 29 rAAV4 1267WO2016115382A1 SEQ ID NO: 30 rAAV4 1268 WO2016115382A1 SEQ ID NO: 31rAAV4 1269 WO2016115382A1 SEQ ID NO: 32 rAAV4 1270 WO2016115382A1 SEQ IDNO: 33 AAV2/8 1271 WO2016131981A1 SEQ ID NO: 47 AAV2/8 1272WO2016131981A1 SEQ ID NO: 48 ancestral AAV 1273 WO2016154344A1 SEQ IDNO: 7 ancestral AAV variant C4 1274 WO2016154344A1 SEQ ID NO: 13ancestral AAV variant C7 1275 WO2016154344A1 SEQ ID NO: 14 ancestral AAVvariant G4 1276 WO2016154344A1 SEQ ID NO: 15 consensus amino acid 1277WO2016154344A1 SEQ ID NO: 16 sequence of ancestral AAV variants, C4, C7and G4 consensus amino acid 1278 WO2016154344A1 SEQ ID NO: 17 sequenceof ancestral AAV variants, C4 and C7 AAV8 (with a AAV2 1279WO2016150403A1 SEQ ID NO: 13 phospholipase domain) AAV VR-942n 1280US20160289275A1 SEQ ID NO: 10 AAV5-A (M569V) 1281 US20160289275A1 SEQ IDNO: 13 AAV5-A (M569V) 1282 US20160289275A1 SEQ ID NO: 14 AAV5-A (Y585V)1283 US20160289275A1 SEQ ID NO: 16 AAV5-A (Y585V) 1284 US20160289275A1SEQ ID NO: 17 AAV5-A (L587T) 1285 US20160289275A1 SEQ ID NO: 19 AAV5-A(L587T) 1286 US20160289275A1 SEQ ID NO: 20 AAV5-A (Y585V/L587T) 1287US20160289275A1 SEQ ID NO: 22 AAV5-A (Y585V/L587T) 1288 US20160289275A1SEQ ID NO: 23 AAV5-B (D652A) 1289 US20160289275A1 SEQ ID NO: 25 AAV5-B(D652A) 1290 US20160289275A1 SEQ ID NO: 26 AAV5-B (T362M) 1291US20160289275A1 SEQ ID NO: 28 AAV5-B (T362M) 1292 US20160289275A1 SEQ IDNO: 29 AAV5-B (Q359D) 1293 US20160289275A1 SEQ ID NO: 31 AAV5-B (Q359D)1294 US20160289275A1 SEQ ID NO: 32 AAV5-B (E350Q) 1295 US20160289275A1SEQ ID NO: 34 AAV5-B (E350Q) 1296 US20160289275A1 SEQ ID NO: 35 AAV5-B(P533S) 1297 US20160289275A1 SEQ ID NO: 37 AAV5-B (P533S) 1298US20160289275A1 SEQ ID NO: 38 AAV5-B (P533G) 1299 US20160289275A1 SEQ IDNO: 40 AAV5-B (P533G) 1300 US20160289275A1 SEQ ID NO: 41 AAV5-mutationin loop VII 1301 US20160289275A1 SEQ ID NO: 43 AAV5-mutation in loop VII1302 US20160289275A1 SEQ ID NO: 44 AAV8 1303 US20160289275A1 SEQ ID NO:47 Mut A (LK03/AAV8) 1304 WO2016181123A1 SEQ ID NO: 1 Mut B (LK03/AAV5)1305 WO2016181123A1 SEQ ID NO: 2 Mut C (AAV8/AAV3B) 1306 WO2016181123A1SEQ ID NO: 3 Mut D (AAV5/AAV3B) 1307 WO2016181123A1 SEQ ID NO: 4 Mut E(AAV8/AAV3B) 1308 WO2016181123A1 SEQ ID NO: 5 Mut F (AAV3B/AAV8) 1309WO2016181123A1 SEQ ID NO: 6 AAV44.9 1310 WO2016183297A1 SEQ ID NO: 4AAV44.9 1311 WO2016183297A1 SEQ ID NO: 5 AAVrh8 1312 WO2016183297A1 SEQID NO: 6 AAV44.9 (S470N) 1313 WO2016183297A1 SEQ ID NO: 9 rh74 VP1 1314US20160375110A1 SEQ ID NO: 1 AAV-LK03 (L125I) 1315 WO2017015102A1 SEQ IDNO: 5 AAV3B (S663V + T492V) 1316 WO2017015102A1 SEQ ID NO: 6 Anc80 1317WO2017019994A2 SEQ ID NO: 1 Anc80 1318 WO2017019994A2 SEQ ID NO: 2 Anc811319 WO2017019994A2 SEQ ID NO: 3 Anc81 1320 WO2017019994A2 SEQ ID NO: 4Anc82 1321 WO2017019994A2 SEQ ID NO: 5 Anc82 1322 WO2017019994A2 SEQ IDNO: 6 Anc83 1323 WO2017019994A2 SEQ ID NO: 7 Anc83 1324 WO2017019994A2SEQ ID NO: 8 Anc84 1325 WO2017019994A2 SEQ ID NO: 9 Anc84 1326WO2017019994A2 SEQ ID NO: 10 Anc94 1327 WO2017019994A2 SEQ ID NO: 11Anc94 1328 WO2017019994A2 SEQ ID NO: 12 Anc113 1329 WO2017019994A2 SEQID NO: 13 Anc113 1330 WO2017019994A2 SEQ ID NO: 14 Anc126 1331WO2017019994A2 SEQ ID NO: 15 Anc126 1332 WO2017019994A2 SEQ ID NO: 16Anc127 1333 WO2017019994A2 SEQ ID NO: 17 Anc127 1334 WO2017019994A2 SEQID NO: 18 Anc80L27 1335 WO2017019994A2 SEQ ID NO: 19 Anc80L59 1336WO2017019994A2 SEQ ID NO: 20 Anc80L60 1337 WO2017019994A2 SEQ ID NO: 21Anc80L62 1338 WO2017019994A2 SEQ ID NO: 22 Anc80L65 1339 WO2017019994A2SEQ ID NO: 23 Anc80L33 1340 WO2017019994A2 SEQ ID NO: 24 Anc80L36 1341WO2017019994A2 SEQ ID NO: 25 Anc80L44 1342 WO2017019994A2 SEQ ID NO: 26Anc80L1 1343 WO2017019994A2 SEQ ID NO: 35 Anc80L1 1344 WO2017019994A2SEQ ID NO: 36 AAVrh10 1345 WO2017019994A2 SEQ ID NO: 41 Anc110 1346WO2017019994A2 SEQ ID NO: 42 Anc110 1347 WO2017019994A2 SEQ ID NO: 43AAVrh32.33 1348 WO2017019994A2 SEQ ID NO: 45 AAVrh74 1349 WO2017049031A1SEQ ID NO: 1 AAV2 1350 WO2017053629A2 SEQ ID NO: 49 AAV2 1351WO2017053629A2 SEQ ID NO: 50 AAV2 1352 WO2017053629A2 SEQ ID NO: 82Parvo-like virus 1353 WO2017070476A2 SEQ ID NO: 1 Parvo-like virus 1354WO2017070476A2 SEQ ID NO: 2 Parvo-like virus 1355 WO2017070476A2 SEQ IDNO: 3 Parvo-like virus 1356 WO2017070476A2 SEQ ID NO: 4 Parvo-like virus1357 WO2017070476A2 SEQ ID NO: 5 Parvo-like virus 1358 WO2017070476A2SEQ ID NO: 6 AAVrh.10 1359 WO2017070516A1 SEQ ID NO: 7 AAVrh.10 1360WO2017070516A1 SEQ ID NO: 14 AAV2tYF 1361 WO2017070491A1 SEQ ID NO: 1AAV-SPK 1362 WO2017075619A1 SEQ ID NO: 28 AAV2.5 1363 US20170128528A1SEQ ID NO: 13 AAV1.1 1364 US20170128528A1 SEQ ID NO: 15 AAV6.1 1365US20170128528A1 SEQ ID NO: 17 AAV6.3.1 1366 US20170128528A1 SEQ ID NO:18 AAV2i8 1367 US20170128528A1 SEQ ID NO: 28 AAV2i8 1368 US20170128528A1SEQ ID NO: 29 ttAAV 1369 US20170128528A1 SEQ ID NO: 30 ttAAV-S312N 1370US20170128528A1 SEQ ID NO: 32 ttAAV-S312N 1371 US20170128528A1 SEQ IDNO: 33 AAV6 (Y705, Y731, and 1372 WO2016134337A1 SEQ ID NO: 24 T492)AAV2 1373 WO2016134375A1 SEQ ID NO: 9 AAV2 1374 WO2016134375A1 SEQ IDNO: 10

In one embodiment, the AAV serotype may be, or may have a sequence asdescribed in International Patent Publication WO2015038958, the contentsof which are herein incorporated by reference in their entirety, suchas, but not limited to, AAV9 (SEQ ID NO: 2 and 11 of WO2015038958 or SEQID NO: 132 and 131 respectively herein), PHP.B (SEQ ID NO: 8 and 9 ofWO2015038958 or SEQ ID NO: 1 and 2 herein), G2B-13 (SEQ ID NO: 12 ofWO2015038958 or SEQ ID NO: 3 herein), G2B-26 (SEQ ID NO: 13 ofWO2015038958 or SEQ ID NO: 1 herein), TH1.1-32 (SEQ ID NO: 14 ofWO2015038958 or SEQ ID NO: 4 herein), TH1.1-35 (SEQ ID NO: 15 ofWO2015038958 or SEQ ID NO: 5 herein) or variants thereof. Further, anyof the targeting peptides or amino acid inserts described inWO2015038958, may be inserted into any parent AAV serotype, such as, butnot limited to, AAV9 (SEQ ID NO: 131 for the DNA sequence and SEQ ID NO:132 for the amino acid sequence). In one embodiment, the amino acidinsert is inserted between amino acids 586-592 of the parent AAV (e.g.,AAV9). In another embodiment, the amino acid insert is inserted betweenamino acids 588-589 of the parent AAV sequence. The amino acid insertmay be, but is not limited to, any of the following amino acidsequences, TLAVPFK (SEQ ID NO: 1 of WO2015038958; herein SEQ ID NO:876), KFPVALT (SEQ ID NO: 3 of WO2015038958; herein SEQ ID NO: 877),LAVPFK (SEQ ID NO: 31 of WO2015038958; herein SEQ ID NO: 878), AVPFK(SEQ ID NO: 32 of WO2015038958; herein SEQ ID NO: 879), VPFK (SEQ ID NO:33 of WO2015038958; herein SEQ ID NO: 880), TLAVPF (SEQ ID NO: 34 ofWO2015038958; herein SEQ ID NO: 881), TLAVP (SEQ ID NO: 35 ofWO2015038958; herein SEQ ID NO: 882), TLAV (SEQ ID NO: 36 ofWO2015038958; herein SEQ ID NO: 883), SVSKPFL (SEQ ID NO: 28 ofWO2015038958; herein SEQ ID NO: 884), FTLTTPK (SEQ ID NO: 29 ofWO2015038958; herein SEQ ID NO: 885), MNATKNV (SEQ ID NO: 30 ofWO2015038958; herein SEQ ID NO: 886), QSSQTPR (SEQ ID NO: 54 ofWO2015038958; herein SEQ ID NO: 887), ILGTGTS (SEQ ID NO: 55 ofWO2015038958; herein SEQ ID NO: 888), TRTNPEA (SEQ ID NO: 56 ofWO2015038958; herein SEQ ID NO: 889), NGGTSSS (SEQ ID NO: 58 ofWO2015038958; herein SEQ ID NO: 890), or YTLSQGW (SEQ ID NO: 60 ofWO2015038958; herein SEQ ID NO: 891). Non-limiting examples ofnucleotide sequences that may encode the amino acid inserts include thefollowing, AAGTTTCCTGTGGCGTTGACT (for SEQ ID NO: 3 of WO2015038958;herein SEQ ID NO: 892), ACTTTGGCGGTGCCTTTTAAG (SEQ ID NO: 24 and 49 ofWO2015038958; herein SEQ ID NO: 893), AGTGTGAGTAAGCCTTTTTTG (SEQ ID NO:25 of WO2015038958; herein SEQ ID NO: 894), TTTACGTTGACGACGCCTAAG (SEQID NO: 26 of WO2015038958; herein SEQ ID NO: 895), ATGAATGCTACGAAGAATGTG(SEQ ID NO: 27 of WO2015038958; herein SEQ ID NO: 896),CAGTCGTCGCAGACGCCTAGG (SEQ ID NO: 48 of WO2015038958; herein SEQ ID NO:897), ATTCTGGGGACTGGTACTTCG (SEQ ID NO: 50 and 52 of WO2015038958;herein SEQ ID NO: 898), ACGCGGACTAATCCTGAGGCT (SEQ ID NO: 51 ofWO2015038958; herein SEQ ID NO: 899), AATGGGGGGACTAGTAGTTCT (SEQ ID NO:53 of WO2015038958; herein SEQ ID NO: 900), or TATACTTTGTCGCAGGGTTGG(SEQ ID NO: 59 of WO2015038958; herein SEQ ID NO: 901).

In one embodiment, the AAV serotype may be, or may have a sequence asdescribed in International Patent Publication WO2017100671, the contentsof which are herein incorporated by reference in their entirety, suchas, but not limited to, AAV9 (SEQ ID NO: 45 of WO2017100671, herein SEQID NO: 875), PHP.N (SEQ ID NO: 46 of WO2017100671, herein SEQ ID NO:873), PHP.S (SEQ ID NO: 47 of WO2017100671, herein SEQ ID NO: 874), orvariants thereof. Further, any of the targeting peptides or amino acidinserts described in WO2017100671 may be inserted into any parent AAVserotype, such as, but not limited to, AAV9 (SEQ ID NO: 127 or SEQ IDNO: 875). In one embodiment, the amino acid insert is inserted betweenamino acids 586-592 of the parent AAV (e.g., AAV9). In anotherembodiment, the amino acid insert is inserted between amino acids588-589 of the parent AAV sequence. The amino acid insert may be, but isnot limited to, any of the following amino acid sequences, AQTLAVPFKAQ(SEQ ID NO: 1 of WO2017100671; herein SEQ ID NO: 902), AQSVSKPFLAQ (SEQID NO: 2 of WO2017100671; herein SEQ ID NO: 903), AQFTLTTPKAQ (SEQ IDNO: 3 in the sequence listing of WO2017100671; herein SEQ ID NO: 904),DGTLAVPFKAQ (SEQ ID NO: 4 in the sequence listing of WO2017100671;herein SEQ ID NO: 905), ESTLAVPFKAQ (SEQ ID NO: 5 of WO2017100671;herein SEQ ID NO: 906), GGTLAVPFKAQ (SEQ ID NO: 6 of WO2017100671;herein SEQ ID NO: 907), AQTLATPFKAQ (SEQ ID NO: 7 and 33 ofWO2017100671; herein SEQ ID NO: 908), ATTLATPFKAQ (SEQ ID NO: 8 ofWO2017100671; herein SEQ ID NO: 909), DGTLATPFKAQ (SEQ ID NO: 9 ofWO2017100671; herein SEQ ID NO: 910), GGTLATPFKAQ (SEQ ID NO: 10 ofWO2017100671; herein SEQ ID NO: 911), SGSLAVPFKAQ (SEQ ID NO: 11 ofWO2017100671; herein SEQ ID NO: 912), AQTLAQPFKAQ (SEQ ID NO: 12 ofWO2017100671; herein SEQ ID NO: 913), AQTLQQPFKAQ (SEQ ID NO: 13 ofWO2017100671; herein SEQ ID NO: 914), AQTLSNPFKAQ (SEQ ID NO: 14 ofWO2017100671; herein SEQ ID NO: 915), AQTLAVPFSNP (SEQ ID NO: 15 ofWO2017100671; herein SEQ ID NO: 916), QGTLAVPFKAQ (SEQ ID NO: 16 ofWO2017100671; herein SEQ ID NO: 917), NQTLAVPFKAQ (SEQ ID NO: 17 ofWO2017100671; herein SEQ ID NO: 918), EGSLAVPFKAQ (SEQ ID NO: 18 ofWO2017100671; herein SEQ ID NO: 919), SGNLAVPFKAQ (SEQ ID NO: 19 ofWO2017100671; herein SEQ ID NO: 920), EGTLAVPFKAQ (SEQ ID NO: 20 ofWO2017100671; herein SEQ ID NO: 921), DSTLAVPFKAQ (SEQ ID NO: 21 inTable 1 of WO2017100671; herein SEQ ID NO: 922), AVTLAVPFKAQ (SEQ ID NO:22 of WO2017100671; herein SEQ ID NO: 923), AQTLSTPFKAQ (SEQ ID NO: 23of WO2017100671; herein SEQ ID NO: 924), AQTLPQPFKAQ (SEQ ID NO: 24 and32 of WO2017100671; herein SEQ ID NO: 925), AQTLSQPFKAQ (SEQ ID NO: 25of WO2017100671; herein SEQ ID NO: 926), AQTLQLPFKAQ (SEQ ID NO: 26 ofWO2017100671; herein SEQ ID NO: 927), AQTLTMPFKAQ (SEQ ID NO: 27, and 34of WO2017100671 and SEQ ID NO: 35 in the sequence listing ofWO2017100671; herein SEQ ID NO: 928), AQTLTTPFKAQ (SEQ ID NO: 28 ofWO2017100671; herein SEQ ID NO: 929), AQYTLSQGWAQ (SEQ ID NO: 29 ofWO2017100671; herein SEQ ID NO: 930), AQMNATKNVAQ (SEQ ID NO: 30 ofWO2017100671; herein SEQ ID NO: 931), AQVSGGHHSAQ (SEQ ID NO: 31 ofWO2017100671; herein SEQ ID NO: 932), AQTLTAPFKAQ (SEQ ID NO: 35 inTable 1 of WO2017100671; herein SEQ ID NO: 933), AQTLSKPFKAQ (SEQ ID NO:36 of WO2017100671; herein SEQ ID NO: 934), QAVRTSL (SEQ ID NO: 37 ofWO2017100671; herein SEQ ID NO: 935), YTLSQGW (SEQ ID NO: 38 ofWO2017100671; herein SEQ ID NO: 891), LAKERLS (SEQ ID NO: 39 ofWO2017100671; herein SEQ ID NO: 936), TLAVPFK (SEQ ID NO: 40 in thesequence listing of WO2017100671; herein SEQ ID NO: 876), SVSKPFL (SEQID NO: 41 of WO2017100671; herein SEQ ID NO: 884), FTLTTPK (SEQ ID NO:42 of WO2017100671; herein SEQ ID NO: 885), MNSTKNV (SEQ ID NO: 43 ofWO2017100671; herein SEQ ID NO: 937), VSGGHHS (SEQ ID NO: 44 ofWO2017100671; herein SEQ ID NO: 938), SAQTLAVPFKAQAQ (SEQ ID NO: 48 ofWO2017100671; herein SEQ ID NO: 939), SXXXLAVPFKAQAQ (SEQ ID NO: 49 ofWO2017100671 wherein X may be any amino acid; herein SEQ ID NO: 940),SAQXXXVPFKAQAQ (SEQ ID NO: 50 of WO2017100671 wherein X may be any aminoacid; herein SEQ ID NO: 941), SAQTLXXXFKAQAQ (SEQ ID NO: 51 ofWO2017100671 wherein X may be any amino acid; herein SEQ ID NO: 942),SAQTLAVXXXAQAQ (SEQ ID NO: 52 of WO2017100671 wherein X may be any aminoacid; herein SEQ ID NO: 943), SAQTLAVPFXXXAQ (SEQ ID NO: 53 ofWO2017100671 wherein X may be any amino acid; herein SEQ ID NO: 944),TNHQSAQ (SEQ ID NO: 65 of WO2017100671; herein SEQ ID NO: 945), AQAQTGW(SEQ ID NO: 66 of WO2017100671; herein SEQ ID NO: 946), DGTLATPFK (SEQID NO: 67 of WO2017100671; herein SEQ ID NO: 947), DGTLATPFKXX (SEQ IDNO: 68 of WO2017100671 wherein X may be any amino acid; herein SEQ IDNO: 948), LAVPFKAQ (SEQ ID NO: 80 of WO2017100671; herein SEQ ID NO:949), VPFKAQ (SEQ ID NO: 81 of WO2017100671; herein SEQ ID NO: 950),FKAQ (SEQ ID NO: 82 of WO2017100671; herein SEQ ID NO: 951), AQTLAV (SEQID NO: 83 of WO2017100671; herein SEQ ID NO: 952), AQTLAVPF (SEQ ID NO:84 of WO2017100671; herein SEQ ID NO: 953), QAVR (SEQ ID NO: 85 ofWO2017100671; herein SEQ ID NO: 954), AVRT (SEQ ID NO: 86 ofWO2017100671; herein SEQ ID NO: 955), VRTS (SEQ ID NO: 87 ofWO2017100671; herein SEQ ID NO: 956), RTSL (SEQ ID NO: 88 ofWO2017100671; herein SEQ ID NO: 957), QAVRT (SEQ ID NO: 89 ofWO2017100671; herein SEQ ID NO: 958), AVRTS (SEQ ID NO: 90 ofWO2017100671; herein SEQ ID NO: 959), VRTSL (SEQ ID NO: 91 ofWO2017100671; herein SEQ ID NO: 960), QAVRTS (SEQ ID NO: 92 ofWO2017100671; herein SEQ ID NO: 961), or AVRTSL (SEQ ID NO: 93 ofWO2017100671; herein SEQ ID NO: 962).

Non-limiting examples of nucleotide sequences that may encode the aminoacid inserts include the following, GATGGGACTTTGGCGGTGCCTTTTAAGGCACAG(SEQ ID NO: 54 of WO2017100671; herein SEQ ID NO: 963),GATGGGACGTTGGCGGTGCCTTTTAAGGCACAG (SEQ ID NO: 55 of WO2017100671; hereinSEQ ID NO: 964), CAGGCGGTTAGGACGTCTTTG (SEQ ID NO: 56 of WO2017100671;herein SEQ ID NO: 965), CAGGTCTTCACGGACTCAGACTATCAG (SEQ ID NO: 57 and78 of WO2017100671; herein SEQ ID NO: 966),CAAGTAAAACCTCTACAAATGTGGTAAAATCG (SEQ ID NO: 58 of WO2017100671; hereinSEQ ID NO: 967), ACTCATCGACCAATACTTGTACTATCTCTCTAGAAC (SEQ ID NO: 59 ofWO2017100671; herein SEQ ID NO: 968), GGAAGTATTCCTTGGTTTTGAACCCA (SEQ IDNO: 60 of WO2017100671; herein SEQ ID NO: 969), GGTCGCGGTTCTTGTTTGTGGAT(SEQ ID NO: 61 of WO2017100671; herein SEQ ID NO: 970),CGACCTTGAAGCGCATGAACTCCT (SEQ ID NO: 62 of WO2017100671; herein SEQ IDNO: 971), GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGCCTGTGCNMNNTTGGGCACTCTGGTGGTTTGTC (SEQ ID NO: 63 of WO2017100671 wherein N maybe A, C, T, or G; herein SEQ ID NO: 972),GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGC AAAAGGCACCGCCAAA GTTTG (SEQ ID NO: 69of WO2017100671 wherein N may be A, C, T, or G; herein SEQ ID NO: 973),GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGCCTGTGCCACCGCCAAAG TTTGGGCACT (SEQ IDNO: 70 of WO2017100671 wherein N may be A, C, T, or G; herein SEQ ID NO:974), GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGCCTGTGCCTTAACAAAGTTTGGGCACTCTGGTGG (SEQ ID NO: 71 of WO2017100671 wherein N may be A,C, T, or G; herein SEQ ID NO: 975),GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGCCTGTGCCTTAAAAGGCACMNNTTGGGCACTCTGGTGGTTTGTG (SEQ ID NO: 72 of WO2017100671 wherein N maybe A, C, T, or G; herein SEQ ID NO: 976), ACTTTGGCGGTGCCTTTTAAG (SEQ IDNO: 74 of WO2017100671; herein SEQ ID NO: 893), AGTGTGAGTAAGCCTTTTTTG(SEQ ID NO: 75 of WO2017100671; herein SEQ ID NO: 894),TTTACGTTGACGACGCCTAAG (SEQ ID NO: 76 of WO2017100671; herein SEQ ID NO:895), TATACTTTGTCGCAGGGTTGG (SEQ ID NO: 77 of WO2017100671; herein SEQID NO: 901), or CTTGCGAAGGAGCGGCTTTCG (SEQ ID NO: 79 of WO2017100671;herein SEQ ID NO: 977).

In one embodiment, the AAV serotype may be, or may have a sequence asdescribed in U.S. Pat. No. 9,624,274, the contents of which are hereinincorporated by reference in their entirety, such as, but not limitedto, AAV1 (SEQ ID NO: 181 of U.S. Pat. No. 9,624,274), AAV6 (SEQ ID NO:182 of U.S. Pat. No. 9,624,274), AAV2 (SEQ ID NO: 183 of U.S. Pat. No.9,624,274), AAV3b (SEQ ID NO: 184 of U.S. Pat. No. 9,624,274), AAV7 (SEQID NO: 185 of U.S. Pat. No. 9,624,274), AAV8 (SEQ ID NO: 186 of U.S.Pat. No. 9,624,274), AAV10 (SEQ ID NO: 187 of U.S. Pat. No. 9,624,274),AAV4 (SEQ ID NO: 188 of U.S. Pat. No. 9,624,274), AAV11 (SEQ ID NO: 189of U.S. Pat. No. 9,624,274), bAAV (SEQ ID NO: 190 of U.S. Pat. No.9,624,274), AAV5 (SEQ ID NO: 191 of U.S. Pat. No. 9,624,274), GPV (SEQID NO: 192 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 992), B19 (SEQID NO: 193 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 993), MVM (SEQID NO: 194 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 994), FPV (SEQID NO: 195 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 995), CPV (SEQID NO: 196 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 996) orvariants thereof. Further, any of the structural protein insertsdescribed in U.S. Pat. No. 9,624,274, may be inserted into, but notlimited to, 1-453 and 1-587 of any parent AAV serotype, such as, but notlimited to, AAV2 (SEQ ID NO: 183 of U.S. Pat. No. 9,624,274). The aminoacid insert may be, but is not limited to, any of the following aminoacid sequences, VNLTWSRASG (SEQ ID NO: 50 of U.S. Pat. No. 9,624,274;herein SEQ ID NO: 1375), EFCINHRGYWVCGD (SEQ ID NO:55 of U.S. Pat. No.9,624,274; herein SEQ ID NO: 1376), EDGQVMDVDLS (SEQ ID NO: 85 of U.S.Pat. No. 9,624,274; herein SEQ ID NO: 1377), EKQRNGTLT (SEQ ID NO: 86 ofU.S. Pat. No. 9,624,274; herein SEQ ID NO: 1378), TYQCRVTHPHLPRALMR (SEQID NO: 87 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1379),RHSTTQPRKTKGSG (SEQ ID NO: 88 of U.S. Pat. No. 9,624,274; herein SEQ IDNO: 1380), DSNPRGVSAYLSR (SEQ ID NO: 89 of U.S. Pat. No. 9,624,274;herein SEQ ID NO: 1381), TITCLWDLAPSK (SEQ ID NO: 90 of U.S. Pat. No.9,624,274; herein SEQ ID NO: 1382), KTKGSGFFVF (SEQ ID NO: 91 of U.S.Pat. No. 9,624,274; herein SEQ ID NO: 1383), THPHLPRALMRS (SEQ ID NO: 92of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1384),GETYQCRVTHPHLPRALMRSTTK (SEQ ID NO: 93 of U.S. Pat. No. 9,624,274;herein SEQ ID NO: 1385), LPRALMRS (SEQ ID NO: 94 of U.S. Pat. No.9,624,274; herein SEQ ID NO: 1386), INHRGYWV (SEQ ID NO: 95 of U.S. Pat.No. 9,624,274; herein SEQ ID NO: 1387), CDAGSVRTNAPD (SEQ ID NO: 60 ofU.S. Pat. No. 9,624,274; herein SEQ ID NO: 1388), AKAVSNLTESRSESLQS (SEQID NO: 96 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1389),SLTGDEFKKVLET (SEQ ID NO: 97 of U.S. Pat. No. 9,624,274; herein SEQ IDNO: 1390), REAVAYRFEED (SEQ ID NO: 98 of U.S. Pat. No. 9,624,274; hereinSEQ ID NO: 1391), INPEIITLDG (SEQ ID NO: 99 of U.S. Pat. No. 9,624,274;herein SEQ ID NO: 1392), DISVTGAPVITATYL (SEQ ID NO: 100 of U.S. Pat.No. 9,624,274; herein SEQ ID NO: 1393), DISVTGAPVITA (SEQ ID NO: 101 ofU.S. Pat. No. 9,624,274; herein SEQ ID NO: 1394), PKTVSNLTESSSESVQS (SEQID NO: 102 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1395),SLMGDEFKAVLET (SEQ ID NO: 103 of U.S. Pat. No. 9,624,274; herein SEQ IDNO: 1396), QHSVAYTFEED (SEQ ID NO: 104 of U.S. Pat. No. 9,624,274;herein SEQ ID NO: 1397), INPEIITRDG (SEQ ID NO: 105 of U.S. Pat. No.9,624,274; herein SEQ ID NO: 1398), DISLTGDPVITASYL (SEQ ID NO: 106 ofU.S. Pat. No. 9,624,274; herein SEQ ID NO: 1399), DISLTGDPVITA (SEQ IDNO: 107 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1400), DQSIDFEIDSA(SEQ ID NO: 108 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1401),KNVSEDLPLPTFSPTLLGDS (SEQ ID NO: 109 of U.S. Pat. No. 9,624,274; hereinSEQ ID NO: 1402), KNVSEDLPLPT (SEQ ID NO: 110 of U.S. Pat. No.9,624,274; herein SEQ ID NO: 1403), CDSGRVRTDAPD (SEQ ID NO: 111 of U.S.Pat. No. 9,624,274; herein SEQ ID NO: 1404), FPEHLLVDFLQSLS (SEQ ID NO:112 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1405), DAEFRHDSG (SEQID NO: 65 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1406),HYAAAQWDFGNTMCQL (SEQ ID NO: 113 of U.S. Pat. No. 9,624,274; herein SEQID NO: 1407), YAAQWDFGNTMCQ (SEQ ID NO: 114 of U.S. Pat. No. 9,624,274;herein SEQ ID NO: 1408), RSQKEGLHYT (SEQ ID NO: 115 of U.S. Pat. No.9,624,274; herein SEQ ID NO: 1409), SSRTPSDKPVAHWANPQAE (SEQ ID NO: 116of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1410), SRTPSDKPVAHWANP(SEQ ID NO: 117 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1411),SSRTPSDKP (SEQ ID NO: 118 of U.S. Pat. No. 9,624,274; herein SEQ ID NO:1412), NADGNVDYHIVINSVP (SEQ ID NO: 119 of U.S. Pat. No. 9,624,274;herein SEQ ID NO: 1413), DGNVDYHMNSV (SEQ ID NO: 120 of U.S. Pat. No.9,624,274; herein SEQ ID NO: 1414), RSFKEFLQSSLRALRQ (SEQ ID NO: 121 ofU.S. Pat. No. 9,624,274; herein SEQ ID NO: 1415); FKEFLQSSLRA (SEQ IDNO: 122 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1416), orQMWAPQWGPD (SEQ ID NO: 123 of U.S. Pat. No. 9,624,274; herein SEQ ID NO:1417).

In one embodiment, the AAV serotype may be, or may have a sequence asdescribed in U.S. Pat. No. 9,475,845, the contents of which are hereinincorporated by reference in their entirety, such as, but not limitedto, AAV capsid proteins comprising modification of one or more aminoacids at amino acid positions 585 to 590 of the native AAV2 capsidprotein. Further the modification may result in, but not limited to, theamino acid sequence RGNRQA (SEQ ID NO: 3 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1418), SSSTDP (SEQ ID NO: 4 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1419), SSNTAP (SEQ ID NO: 5 of U.S. Pat.No. 9,475,845; herein SEQ ID NO: 1420), SNSNLP (SEQ ID NO: 6 of U.S.Pat. No. 9,475,845; herein SEQ ID NO: 1421), SSTTAP (SEQ ID NO: 7 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1422), AANTAA (SEQ ID NO: 8of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1423), QQNTAP (SEQ ID NO:9 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1424), SAQAQA (SEQ IDNO: 10 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1425), QANTGP (SEQID NO: 11 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1426), NATTAP(SEQ ID NO: 12 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1427),SSTAGP (SEQ ID NO: 13 and 20 of U.S. Pat. No. 9,475,845; herein SEQ IDNO: 1428), QQNTAA (SEQ ID NO: 14 of U.S. Pat. No. 9,475,845; herein SEQID NO: 1429), PSTAGP (SEQ ID NO: 15 of U.S. Pat. No. 9,475,845; hereinSEQ ID NO: 1430), NQNTAP (SEQ ID NO: 16 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1431), QAANAP (SEQ ID NO: 17 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1432), SIVGLP (SEQ ID NO: 18 of U.S. Pat.No. 9,475,845; herein SEQ ID NO: 1433), AASTAA (SEQ ID NO: 19, and 27 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1434), SQNTTA (SEQ ID NO: 21of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1435), QQDTAP (SEQ ID NO:22 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1436), QTNTGP (SEQ IDNO: 23 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1437), QTNGAP (SEQID NO: 24 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1438), QQNAAP(SEQ ID NO: 25 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1439), orAANTQA (SEQ ID NO: 26 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:1440). In one embodiment, the amino acid modification is a substitutionat amino acid positions 262 through 265 in the native AAV2 capsidprotein or the corresponding position in the capsid protein of anotherAAV with a targeting sequence. The targeting sequence may be, but is notlimited to, any of the amino acid sequences, NGRAHA (SEQ ID NO: 38 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1441), QPEHSST (SEQ ID NO: 39and 50 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1442), VNTANST (SEQID NO: 40 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1443), HGPMQKS(SEQ ID NO: 41 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1444),PHKPPLA (SEQ ID NO: 42 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:1445), IKNNEMW (SEQ ID NO: 43 of U.S. Pat. No. 9,475,845; herein SEQ IDNO: 1446), RNLDTPM (SEQ ID NO: 44 of U.S. Pat. No. 9,475,845; herein SEQID NO: 1447), VDSHRQS (SEQ ID NO: 45 of U.S. Pat. No. 9,475,845; hereinSEQ ID NO: 1448), YDSKTKT (SEQ ID NO: 46 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1449), SQLPHQK (SEQ ID NO: 47 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1450), STMQQNT (SEQ ID NO: 48 of U.S. Pat.No. 9,475,845; herein SEQ ID NO: 1451), TERYMTQ (SEQ ID NO: 49 of U.S.Pat. No. 9,475,845; herein SEQ ID NO: 1452), DASLSTS (SEQ ID NO: 51 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1453), DLPNKKT (SEQ ID NO: 52of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1454), DLTAARL (SEQ ID NO:53 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1455), EPHQFNY (SEQ IDNO: 54 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1456), EPQSNHT (SEQID NO: 55 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1457), MSSWPSQ(SEQ ID NO: 56 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1458),NPKHNAT (SEQ ID NO: 57 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:1459), PDGMRTT (SEQ ID NO: 58 of U.S. Pat. No. 9,475,845; herein SEQ IDNO: 1460), PNNNKTT (SEQ ID NO: 59 of U.S. Pat. No. 9,475,845; herein SEQID NO: 1461), QSTTHDS (SEQ ID NO: 60 of U.S. Pat. No. 9,475,845; hereinSEQ ID NO: 1462), TGSKQKQ (SEQ ID NO: 61 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1463), SLKHQAL (SEQ ID NO: 62 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1464), SPIDGEQ (SEQ ID NO: 63 of U.S. Pat.No. 9,475,845; herein SEQ ID NO: 1465), WIFPWIQL (SEQ ID NO: 64 and 112of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1466), CDCRGDCFC (SEQ IDNO: 65 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1467), CNGRC (SEQID NO: 66 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1468), CPRECES(SEQ ID NO: 67 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1469),CTTHWGFTLC (SEQ ID NO: 68 and 123 of U.S. Pat. No. 9,475,845; herein SEQID NO: 1470), CGRRAGGSC (SEQ ID NO: 69 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1471), CKGGRAKDC (SEQ ID NO: 70 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1472), CVPELGHEC (SEQ ID NO: 71 and 115 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1473), CRRETAWAK (SEQ ID NO:72 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1474), VSWFSHRYSPFAVS(SEQ ID NO: 73 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1475),GYRDGYAGPILYN (SEQ ID NO: 74 of U.S. Pat. No. 9,475,845; herein SEQ IDNO: 1476), XXXYXXX (SEQ ID NO: 75 of U.S. Pat. No. 9,475,845; herein SEQID NO: 1477), YXNW (SEQ ID NO: 76 of U.S. Pat. No. 9,475,845; herein SEQID NO: 1478), RPLPPLP (SEQ ID NO: 77 of U.S. Pat. No. 9,475,845; hereinSEQ ID NO: 1479), APPLPPR (SEQ ID NO: 78 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1480), DVFYPYPYASGS (SEQ ID NO: 79 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1481), MYWYPY (SEQ ID NO: 80 of U.S. Pat.No. 9,475,845; herein SEQ ID NO: 1482), DITWDQLWDLMK (SEQ ID NO: 81 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1483), CWDDXWLC (SEQ ID NO:82 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1484), EWCEYLGGYLRCYA(SEQ ID NO: 83 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1485),YXCXXGPXTWXCXP (SEQ ID NO: 84 of U.S. Pat. No. 9,475,845; herein SEQ IDNO: 1486), IEGPTLRQWLAARA (SEQ ID NO: 85 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1487), LWXXX (SEQ ID NO: 86 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1488), XFXXYLW (SEQ ID NO: 87 of U.S. Pat.No. 9,475,845; herein SEQ ID NO: 1489), SSIISHFRWGLCD (SEQ ID NO: 88 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1490), MSRPACPPNDKYE (SEQ IDNO: 89 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1491), CLRSGRGC(SEQ ID NO: 90 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1492),CHWMFSPWC (SEQ ID NO: 91 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:1493), WXXF (SEQ ID NO: 92 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:1494), CSSRLDAC (SEQ ID NO: 93 of U.S. Pat. No. 9,475,845; herein SEQ IDNO: 1495), CLPVASC (SEQ ID NO: 94 of U.S. Pat. No. 9,475,845; herein SEQID NO: 1496), CGFECVRQCPERC (SEQ ID NO: 95 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1497), CVALCREACGEGC (SEQ ID NO: 96 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1498), SWCEPGWCR (SEQ ID NO: 97 of U.S.Pat. No. 9,475,845; herein SEQ ID NO: 1499), YSGKWGW (SEQ ID NO: 98 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1500), GLSGGRS (SEQ ID NO: 99of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1501), LMLPRAD (SEQ ID NO:100 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1502), CSCFRDVCC (SEQID NO: 101 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1503),CRDVVSVIC (SEQ ID NO: 102 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:1504), MARSGL (SEQ ID NO: 103 of U.S. Pat. No. 9,475,845; herein SEQ IDNO: 1505), MARAKE (SEQ ID NO: 104 of U.S. Pat. No. 9,475,845; herein SEQID NO: 1506), MSRTMS (SEQ ID NO: 105 of U.S. Pat. No. 9,475,845; hereinSEQ ID NO: 1507), KCCYSL (SEQ ID NO: 106 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1508), MYWGDSHWLQYWYE (SEQ ID NO: 107 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1509), MQLPLAT (SEQ ID NO: 108 of U.S. Pat.No. 9,475,845; herein SEQ ID NO: 1510), EWLS (SEQ ID NO: 109 of U.S.Pat. No. 9,475,845; herein SEQ ID NO: 1511), SNEW (SEQ ID NO: 110 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1512), TNYL (SEQ ID NO: 111of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1513), WDLAWMFRLPVG (SEQID NO: 113 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1514),CTVALPGGYVRVC (SEQ ID NO: 114 of U.S. Pat. No. 9,475,845; herein SEQ IDNO: 1515), CVAYCIEHHCWTC (SEQ ID NO: 116 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1516), CVFAHNYDYLVC (SEQ ID NO: 117 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1517), CVFTSNYAFC (SEQ ID NO: 118 of U.S.Pat. No. 9,475,845; herein SEQ ID NO: 1518), VHSPNKK (SEQ ID NO: 119 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1519), CRGDGWC (SEQ ID NO:120 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1520), XRGCDX (SEQ IDNO: 121 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1521), PXXX (SEQID NO: 122 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1522),SGKGPRQITAL (SEQ ID NO: 124 of U.S. Pat. No. 9,475,845; herein SEQ IDNO: 1523), AAAAAAAAAXXXXX (SEQ ID NO: 125 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1524), VYMSPF (SEQ ID NO: 126 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1525), ATWLPPR (SEQ ID NO: 127 of U.S. Pat.No. 9,475,845; herein SEQ ID NO: 1526), HTMYYHHYQHHL (SEQ ID NO: 128 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1527), SEVGCRAGPLQWLCEKYFG(SEQ ID NO: 129 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1528),CGLLPVGRPDRNVWRWLC (SEQ ID NO: 130 of U.S. Pat. No. 9,475,845; hereinSEQ ID NO: 1529), CKGQCDRFKGLPWEC (SEQ ID NO: 131 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1530), SGRSA (SEQ ID NO: 132 of U.S. Pat.No. 9,475,845; herein SEQ ID NO: 1531), WGFP (SEQ ID NO: 133 of U.S.Pat. No. 9,475,845; herein SEQ ID NO: 1532), AEPMPHSLNFSQYLWYT (SEQ IDNO: 134 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1533), WAYXSP (SEQID NO: 135 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1534), IELLQAR(SEQ ID NO: 136 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1535),AYTKCSRQWRTCMTTH (SEQ ID NO: 137 of U.S. Pat. No. 9,475,845; herein SEQID NO: 1536), PQNSKIPGPTFLDPH (SEQ ID NO: 138 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1537), SMEPALPDWWWKMFK (SEQ ID NO: 139 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1538), ANTPCGPYTHDCPVKR (SEQID NO: 140 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1539),TACHQHVRMVRP (SEQ ID NO: 141 of U.S. Pat. No. 9,475,845; herein SEQ IDNO: 1540), VPWMEPAYQRFL (SEQ ID NO: 142 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1541), DPRATPGS (SEQ ID NO: 143 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1542), FRPNRAQDYNTN (SEQ ID NO: 144 of U.S.Pat. No. 9,475,845; herein SEQ ID NO: 1543), CTKNSYLMC (SEQ ID NO: 145of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1544), CXXTXXXGXGC (SEQ IDNO: 146 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1545), CPIEDRPMC(SEQ ID NO: 147 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1546),HEWSYLAPYPWF (SEQ ID NO: 148 of U.S. Pat. No. 9,475,845; herein SEQ IDNO: 1547), MCPKHPLGC (SEQ ID NO: 149 of U.S. Pat. No. 9,475,845; hereinSEQ ID NO: 1548), RMWPSSTVNLSAGRR (SEQ ID NO: 150 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1549), SAKTAVSQRVWLPSHRGGEP (SEQ ID NO: 151of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1550),KSREHVNNSACPSKRITAAL (SEQ ID NO: 152 of U.S. Pat. No. 9,475,845; hereinSEQ ID NO: 1551), EGFR (SEQ ID NO: 153 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1552), AGLGVR (SEQ ID NO: 154 of U.S. Pat. No.9,475,845; herein SEQ ID NO: 1553), GTRQGHTMRLGVSDG (SEQ ID NO: 155 ofU.S. Pat. No. 9,475,845; herein SEQ ID NO: 1554), IAGLATPGWSHWLAL (SEQID NO: 156 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1555), SMSIARL(SEQ ID NO: 157 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1556),HTFEPGV (SEQ ID NO: 158 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:1557), NTSLKRISNKRIRRK (SEQ ID NO: 159 of US9475845; herein SEQ ID NO:1558), LRIKRKRRKRKKTRK (SEQ ID NO: 160 of U.S. Pat. No. 9,475,845;herein SEQ ID NO: 1559), GGG, GFS, LWS, EGG, LLV, LSP, LBS, AGG, GRR,GGH and GTV.

In one embodiment, the AAV serotype may be, or may have a sequence asdescribed in United States Publication No. US 20160369298, the contentsof which are herein incorporated by reference in their entirety, suchas, but not limited to, site-specific mutated capsid protein of AAV2(SEQ ID NO: 97 of US 20160369298; herein SEQ ID NO: 1560) or variantsthereof, wherein the specific site is at least one site selected fromsites R447, G453, S578, N587, N587+1, S662 of VP1 or fragment thereof.

Further, any of the mutated sequences described in US 20160369298, maybe or may have, but not limited to, any of the following sequencesSDSGASN (SEQ ID NO: 1 and SEQ ID NO: 231 of US20160369298; herein SEQ IDNO: 1561), SPSGASN (SEQ ID NO: 2 of US20160369298; herein SEQ ID NO:1562), SHSGASN (SEQ ID NO: 3 of US20160369298; herein SEQ ID NO: 1563),SRSGASN (SEQ ID NO: 4 of US20160369298; herein SEQ ID NO: 1564), SKSGASN(SEQ ID NO: 5 of US20160369298; herein SEQ ID NO: 1565), SNSGASN (SEQ IDNO: 6 of US20160369298; herein SEQ ID NO: 1566), SGSGASN (SEQ ID NO: 7of US20160369298; herein SEQ ID NO: 1567), SASGASN (SEQ ID NO: 8, 175,and 221 of US20160369298; herein SEQ ID NO: 1568), SESGTSN (SEQ ID NO: 9of US20160369298; herein SEQ ID NO: 1569), STTGGSN (SEQ ID NO: 10 ofUS20160369298; herein SEQ ID NO: 1570), SSAGSTN (SEQ ID NO: 11 ofUS20160369298; herein SEQ ID NO: 1571), NNDSQA (SEQ ID NO: 12 ofUS20160369298; herein SEQ ID NO: 1572), NNRNQA (SEQ ID NO: 13 ofUS20160369298; herein SEQ ID NO: 1573), NNNKQA (SEQ ID NO: 14 ofUS20160369298; herein SEQ ID NO: 1574), NAKRQA (SEQ ID NO: 15 ofUS20160369298; herein SEQ ID NO: 1575), NDEHQA (SEQ ID NO: 16 ofUS20160369298; herein SEQ ID NO: 1576), NTSQKA (SEQ ID NO: 17 ofUS20160369298; herein SEQ ID NO: 1577), YYLSRTNTPSGTDTQSRLVFSQAGA (SEQID NO: 18 of US20160369298; herein SEQ ID NO: 1578),YYLSRTNTDSGTETQSGLDFSQAGA (SEQ ID NO: 19 of US20160369298; herein SEQ IDNO: 1579), YYLSRTNTESGTPTQSALEFSQAGA (SEQ ID NO: 20 of US20160369298;herein SEQ ID NO: 1580), YYLSRTNTHSGTHTQSPLHFSQAGA (SEQ ID NO: 21 ofUS20160369298; herein SEQ ID NO: 1581), YYLSRTNTSSGTITISHLIFSQAGA (SEQID NO: 22 of US20160369298; herein SEQ ID NO: 1582),YYLSRTNTRSGIMTKSSLMFSQAGA (SEQ ID NO: 23 of US20160369298; herein SEQ IDNO: 1583), YYLSRTNTKSGRKTLSNLSFSQAGA (SEQ ID NO: 24 of US20160369298;herein SEQ ID NO: 1584), YYLSRTNDGSGPVTPSKLRFSQRGA (SEQ ID NO: 25 ofUS20160369298; herein SEQ ID NO: 1585), YYLSRTNAASGHATHSDLKFSQPGA (SEQID NO: 26 of US20160369298; herein SEQ ID NO: 1586),YYLSRTNGQAGSLTMSELGFSQVGA (SEQ ID NO: 27 of US20160369298; herein SEQ IDNO: 1587), YYLSRTNSTGGNQTTSQLLFSQLSA (SEQ ID NO: 28 of US20160369298;herein SEQ ID NO: 1588), YFLSRTNNNTGLNTNSTLNFSQGRA (SEQ ID NO: 29 ofUS20160369298; herein SEQ ID NO: 1589), SKTGADNNNSEYSWTG (SEQ ID NO: 30of US20160369298; herein SEQ ID NO: 1590), SKTDADNNNSEYSWTG (SEQ ID NO:31 of US20160369298; herein SEQ ID NO: 1591), SKTEADNNNSEYSWTG (SEQ IDNO: 32 of US20160369298; herein SEQ ID NO: 1592), SKTPADNNNSEYSWTG (SEQID NO: 33 of US20160369298; herein SEQ ID NO: 1593), SKTHADNNNSEYSWTG(SEQ ID NO: 34 of US20160369298; herein SEQ ID NO: 1594),SKTQADNNNSEYSWTG (SEQ ID NO: 35 of US20160369298; herein SEQ ID NO:1595), SKTIADNNNSEYSWTG (SEQ ID NO: 36 of US20160369298; herein SEQ IDNO: 1596), SKTMADNNNSEYSWTG (SEQ ID NO: 37 of US20160369298; herein SEQID NO: 1597), SKTRADNNNSEYSWTG (SEQ ID NO: 38 of US20160369298; hereinSEQ ID NO: 1598), SKTNADNNNSEYSWTG (SEQ ID NO: 39 of US20160369298;herein SEQ ID NO: 1599), SKTVGRNNNSEYSWTG (SEQ ID NO: 40 ofUS20160369298; herein SEQ ID NO: 1600), SKTADRNNNSEYSWTG (SEQ ID NO: 41of US20160369298; herein SEQ ID NO: 1601), SKKLSQNNNSKYSWQG (SEQ ID NO:42 of US20160369298; herein SEQ ID NO: 1602), SKPTTGNNNSDYSWPG (SEQ IDNO: 43 of US20160369298; herein SEQ ID NO: 1603), STQKNENNNSNYSWPG (SEQID NO: 44 of US20160369298; herein SEQ ID NO: 1604), HKDDEGKF (SEQ IDNO: 45 of US20160369298; herein SEQ ID NO: 1605), HKDDNRKF (SEQ ID NO:46 of US20160369298; herein SEQ ID NO: 1606), HKDDTNKF (SEQ ID NO: 47 ofUS20160369298; herein SEQ ID NO: 1607), HEDSDKNF (SEQ ID NO: 48 ofUS20160369298; herein SEQ ID NO: 1608), HRDGADSF (SEQ ID NO: 49 ofUS20160369298; herein SEQ ID NO: 1609), HGDNKSRF (SEQ ID NO: 50 ofUS20160369298; herein SEQ ID NO: 1610), KQGSEKTNVDFEEV (SEQ ID NO: 51 ofUS20160369298; herein SEQ ID NO: 1611), KQGSEKTNVDSEEV (SEQ ID NO: 52 ofUS20160369298; herein SEQ ID NO: 1612), KQGSEKTNVDVEEV (SEQ ID NO: 53 ofUS20160369298; herein SEQ ID NO: 1613), KQGSDKTNVDDAGV (SEQ ID NO: 54 ofUS20160369298; herein SEQ ID NO: 1614), KQGSSKTNVDPREV (SEQ ID NO: 55 ofUS20160369298; herein SEQ ID NO: 1615), KQGSRKTNVDHKQV (SEQ ID NO: 56 ofUS20160369298; herein SEQ ID NO: 1616), KQGSKGGNVDTNRV (SEQ ID NO: 57 ofUS20160369298; herein SEQ ID NO: 1617), KQGSGEANVDNGDV (SEQ ID NO: 58 ofUS20160369298; herein SEQ ID NO: 1618), KQDAAADNIDYDHV (SEQ ID NO: 59 ofUS20160369298; herein SEQ ID NO: 1619), KQSGTRSNAAASSV (SEQ ID NO: 60 ofUS20160369298; herein SEQ ID NO: 1620), KENTNTNDTELTNV (SEQ ID NO: 61 ofUS20160369298; herein SEQ ID NO: 1621), QRGNNVAATADVNT (SEQ ID NO: 62 ofUS20160369298; herein SEQ ID NO: 1622), QRGNNEAATADVNT (SEQ ID NO: 63 ofUS20160369298; herein SEQ ID NO: 1623), QRGNNPAATADVNT (SEQ ID NO: 64 ofUS20160369298; herein SEQ ID NO: 1624), QRGNNHAATADVNT (SEQ ID NO: 65 ofUS20160369298; herein SEQ ID NO: 1625), QEENNIAATPGVNT (SEQ ID NO: 66 ofUS20160369298; herein SEQ ID NO: 1626), QPPNNMAATHEVNT (SEQ ID NO: 67 ofUS20160369298; herein SEQ ID NO: 1627), QHHNNSAATTIVNT (SEQ ID NO: 68 ofUS20160369298; herein SEQ ID NO: 1628), QTTNNRAAFNMVET (SEQ ID NO: 69 ofUS20160369298; herein SEQ ID NO: 1629), QKKNNNAASKKVAT (SEQ ID NO: 70 ofUS20160369298; herein SEQ ID NO: 1630), QGGNNKAADDAVKT (SEQ ID NO: 71 ofUS20160369298; herein SEQ ID NO: 1631), QAAKGGAADDAVKT (SEQ ID NO: 72 ofUS20160369298; herein SEQ ID NO: 1632), QDDRAAAANESVDT (SEQ ID NO: 73 ofUS20160369298; herein SEQ ID NO: 1633), QQQHDDAAYQRVHT (SEQ ID NO: 74 ofUS20160369298; herein SEQ ID NO: 1634), QSSSSLAAVSTVQT (SEQ ID NO: 75 ofUS20160369298; herein SEQ ID NO: 1635), QNNQTTAAIRNVTT (SEQ ID NO: 76 ofUS20160369298; herein SEQ ID NO: 1636), NYNKKSDNVDFT (SEQ ID NO: 77 ofUS20160369298; herein SEQ ID NO: 1637), NYNKKSENVDFT (SEQ ID NO: 78 ofUS20160369298; herein SEQ ID NO: 1638), NYNKKSLNVDFT (SEQ ID NO: 79 ofUS20160369298; herein SEQ ID NO: 1639), NYNKKSPNVDFT (SEQ ID NO: 80 ofUS20160369298; herein SEQ ID NO: 1640), NYSKKSHCVDFT (SEQ ID NO: 81 ofUS20160369298; herein SEQ ID NO: 1641), NYRKTIYVDFT (SEQ ID NO: 82 ofUS20160369298; herein SEQ ID NO: 1642), NYKEKKDVHFT (SEQ ID NO: 83 ofUS20160369298; herein SEQ ID NO: 1643), NYGHRAIVQFT (SEQ ID NO: 84 ofUS20160369298; herein SEQ ID NO: 1644), NYANHQFVVCT (SEQ ID NO: 85 ofUS20160369298; herein SEQ ID NO: 1645), NYDDDPTGVLLT (SEQ ID NO: 86 ofUS20160369298; herein SEQ ID NO: 1646), NYDDPTGVLLT (SEQ ID NO: 87 ofUS20160369298; herein SEQ ID NO: 1647), NFEQQNSVEWT (SEQ ID NO: 88 ofUS20160369298; herein SEQ ID NO: 1648), SQSGASN (SEQ ID NO: 89 and SEQID NO: 241 of US20160369298; herein SEQ ID NO: 1649), NNGSQA (SEQ ID NO:90 of US20160369298; herein SEQ ID NO: 1650), YYLSRTNTPSGTTTWSRLQFSQAGA(SEQ ID NO: 91 of US20160369298; herein SEQ ID NO: 1651),SKTSADNNNSEYSWTG (SEQ ID NO: 92 of US20160369298; herein SEQ ID NO:1652), HKDDEEKF (SEQ ID NO: 93, 209, 214, 219, 224, 234, 239, and 244 ofUS20160369298; herein SEQ ID NO: 1653), KQGSEKTNVDIEEV (SEQ ID NO: 94 ofUS20160369298; herein SEQ ID NO: 1654), QRGNNQAATADVNT (SEQ ID NO: 95 ofUS20160369298; herein SEQ ID NO: 1655), NYNKKSVNVDFT (SEQ ID NO: 96 ofUS20160369298; herein SEQ ID NO: 1656),SQSGASNYNTPSGTTTQSRLQFSTSADNNNSEYSWTGATKYH (SEQ ID NO: 106 ofUS20160369298; herein SEQ ID NO: 1657),SASGASNFNSEGGSLTQSSLGFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 107 ofUS20160369298; herein SEQ ID NO: 1658),SQSGASNYNTPSGTTTQSRLQFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 108 ofUS20160369298; herein SEQ ID NO: 1659),SASGASNYNTPSGTTTQSRLQFSTSADNNNSEFSWPGATTYH (SEQ ID NO: 109 ofUS20160369298; herein SEQ ID NO: 1660),SQSGASNFNSEGGSLTQSSLGFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 110 ofUS20160369298; herein SEQ ID NO: 1661),SASGASNYNTPSGSLTQSSLGFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 111 ofUS20160369298; herein SEQ ID NO: 1662),SQSGASNYNTPSGTTTQSRLQFSTSADNNNSDFSWTGATKYH (SEQ ID NO: 112 ofUS20160369298; herein SEQ ID NO: 1663),SGAGASNFNSEGGSLTQSSLGFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 113 ofUS20160369298; herein SEQ ID NO: 1664), SGAGASN (SEQ ID NO: 176 ofUS20160369298; herein SEQ ID NO: 1665), NSEGGSLTQSSLGFS (SEQ ID NO: 177,185, 193 and 202 of US20160369298; herein SEQ ID NO: 1666), TDGENNNSDFS(SEQ ID NO: 178 of US20160369298; herein SEQ ID NO: 1667), SEFSWPGATT(SEQ ID NO: 179 of US20160369298; herein SEQ ID NO: 1668), TSADNNNSDFSWT(SEQ ID NO: 180 of US20160369298; herein SEQ ID NO: 1669), SQSGASNY (SEQID NO: 181, 187, and 198 of US20160369298; herein SEQ ID NO: 1670),NTPSGTTTQSRLQFS (SEQ ID NO: 182, 188, 191, and 199 of US20160369298;herein SEQ ID NO: 1671), TSADNNNSEYSWTGATKYH (SEQ ID NO: 183 ofUS20160369298; herein SEQ ID NO: 1672), SASGASNF (SEQ ID NO: 184 ofUS20160369298; herein SEQ ID NO: 1673), TDGENNNSDFSWTGATKYH (SEQ ID NO:186, 189, 194, 197, and 203 of US20160369298; herein SEQ ID NO: 1674),SASGASNY (SEQ ID NO: 190 and SEQ ID NO: 195 of US20160369298; herein SEQID NO: 1675), TSADNNNSEFSWPGATTYH (SEQ ID NO: 192 of US20160369298;herein SEQ ID NO: 1676), NTPSGSLTQSSLGFS (SEQ ID NO: 196 ofUS20160369298; herein SEQ ID NO: 1677), TSADNNNSDFSWTGATKYH (SEQ ID NO:200 of US20160369298; herein SEQ ID NO: 1678), SGAGASNF (SEQ ID NO: 201of US20160369298; herein SEQ ID NO: 1679),CTCCAGVVSVVSMRSRVCVNSGCAGCTDHCVVSRNSGTCVMSACACAA (SEQ ID NO: 204 ofUS20160369298; herein SEQ ID NO: 1680),CTCCAGAGAGGCAACAGACAAGCAGCTACCGCAGATGTCAACACACAA (SEQ ID NO: 205 ofUS20160369298; herein SEQ ID NO: 1681), SAAGASN (SEQ ID NO: 206 ofUS20160369298; herein SEQ ID NO: 1682), YFLSRTNTESGSTTQSTLRFSQAG (SEQ IDNO: 207 of US20160369298; herein SEQ ID NO: 1683), SKTSADNNNSDFS (SEQ IDNO: 208, 228, and 253 of US20160369298; herein SEQ ID NO: 1684),KQGSEKTDVDIDKV (SEQ ID NO: 210 of US20160369298; herein SEQ ID NO:1685), STAGASN (SEQ ID NO: 211 of US20160369298; herein SEQ ID NO:1686), YFLSRTNTTSGIETQSTLRFSQAG (SEQ ID NO: 212 and SEQ ID NO: 247 ofUS20160369298; herein SEQ ID NO: 1687), SKTDGENNNSDFS (SEQ ID NO: 213and SEQ ID NO: 248 of US20160369298; herein SEQ ID NO: 1688),KQGAAADDVEIDGV (SEQ ID NO: 215 and SEQ ID NO: 250 of US20160369298;herein SEQ ID NO: 1689), SEAGASN (SEQ ID NO: 216 of US20160369298;herein SEQ ID NO: 1690), YYLSRTNTPSGTTTQSRLQFSQAG (SEQ ID NO: 217, 232and 242 of US20160369298; herein SEQ ID NO: 1691), SKTSADNNNSEYS (SEQ IDNO: 218, 233, 238, and 243 of US20160369298; herein SEQ ID NO: 1692),KQGSEKTNVDIEKV (SEQ ID NO: 220, 225 and 245 of US20160369298; herein SEQID NO: 1693), YFLSRTNDASGSDTKSTLLFSQAG (SEQ ID NO: 222 of US20160369298;herein SEQ ID NO: 1694), STTPSENNNSEYS (SEQ ID NO: 223 of US20160369298;herein SEQ ID NO: 1695), SAAGATN (SEQ ID NO: 226 and SEQ ID NO: 251 ofUS20160369298; herein SEQ ID NO: 1696), YFLSRTNGEAGSATLSELRFSQAG (SEQ IDNO: 227 of US20160369298; herein SEQ ID NO: 1697), HGDDADRF (SEQ ID NO:229 and SEQ ID NO: 254 of US20160369298; herein SEQ ID NO: 1698),KQGAEKSDVEVDRV (SEQ ID NO: 230 and SEQ ID NO: 255 of US20160369298;herein SEQ ID NO: 1699), KQDSGGDNIDIDQV (SEQ ID NO: 235 ofUS20160369298; herein SEQ ID NO: 1700), SDAGASN (SEQ ID NO: 236 ofUS20160369298; herein SEQ ID NO: 1701), YFLSRTNTEGGHDTQSTLRFSQAG (SEQ IDNO: 237 of US20160369298; herein SEQ ID NO: 1702), KEDGGGSDVAIDEV (SEQID NO: 240 of US20160369298; herein SEQ ID NO: 1703), SNAGASN (SEQ IDNO: 246 of US20160369298; herein SEQ ID NO: 1704), andYFLSRTNGEAGSATLSELRFSQPG (SEQ ID NO: 252 of US20160369298; herein SEQ IDNO: 1705). Non-limiting examples of nucleotide sequences that may encodethe amino acid mutated sites include the following,AGCVVMDCAGGARSCASCAAC (SEQ ID NO: 97 of US20160369298; herein SEQ ID NO:1706), AACRACRRSMRSMAGGCA (SEQ ID NO: 98 of US20160369298; herein SEQ IDNO: 1707), CACRRGGACRRCRMSRRSARSTTT (SEQ ID NO: 99 of US20160369298;herein SEQ ID NO: 1708),TATTTCTTGAGCAGAACAAACRVCVVSRSCGGAMNCVHSACGMHSTCAVVSCTTVDSTTTTCTCAGSBCRGSGCG (SEQ ID NO: 100 of US20160369298; herein SEQ ID NO:1709), TCAAMAMMAVNSRVCSRSAACAACAACAGTRASTTCTCGTGGMMAGGA (SEQ ID NO: 101of US20160369298; herein SEQ ID NO: 1710),AAGSAARRCRSCRVSRVARVCRATRYCGMSNHCRVMVRSGTC (SEQ ID NO: 102 ofUS20160369298; herein SEQ ID NO: 1711),CAGVVSVVSMRSRVCVNSGCAGCTDHCVVSRNSGTCVMSACA (SEQ ID NO: 103 ofUS20160369298; herein SEQ ID NO: 1712),AACTWCRVSVASMVSVHSDDTGTGSWSTKSACT (SEQ ID NO: 104 of US20160369298;herein SEQ ID NO: 1713), TTGTTGAACATCACCACGTGACGCACGTTC (SEQ ID NO: 256of US20160369298; herein SEQ ID NO: 1714),TCCCCGTGGTTCTACTACATAATGTGGCCG (SEQ ID NO: 257 of US20160369298; hereinSEQ ID NO: 1715), TTCCACACTCCGTTTTGGATAATGTTGAAC (SEQ ID NO: 258 ofUS20160369298; herein SEQ ID NO: 1716), AGGGACATCCCCAGCTCCATGCTGTGGTCG(SEQ ID NO: 259 of US20160369298; herein SEQ ID NO: 1717),AGGGACAACCCCTCCGACTCGCCCTAATCC (SEQ ID NO: 260 of US20160369298; hereinSEQ ID NO: 1718), TCCTAGTAGAAGACACCCTCTCACTGCCCG (SEQ ID NO: 261 ofUS20160369298; herein SEQ ID NO: 1719), AGTACCATGTACACCCACTCTCCCAGTGCC(SEQ ID NO: 262 of US20160369298; herein SEQ ID NO: 1720),ATATGGACGTTCATGCTGATCACCATACCG (SEQ ID NO: 263 of US20160369298; hereinSEQ ID NO: 1721), AGCAGGAGCTCCTTGGCCTCAGCGTGCGAG (SEQ ID NO: 264 ofUS20160369298; herein SEQ ID NO: 1722), ACAAGCAGCTTCACTATGACAACCACTGAC(SEQ ID NO: 265 of US20160369298; herein SEQ ID NO: 1723),CAGCCTAGGAACTGGCTTCCTGGACCCTGTTACCGCCAGCAGAGAGTCTCAAMAMMAVNSRVCSRSAACAACAACAGTRASTTCTCCTGGMMAGGAGCTACCAAGTACCACCTCAATGGCAGAGACTCTCTGGTGAATCCCGGACCAGCTATGGCAAGCCACRRGGACRRCRMSRRSARSTTTTTTCCTCAGAGCGGGGTTCTCATCTTTGGGAAGSAARRCRSCRVSRVARVCRATRYCGMSNHCRVMVRSGTCATGATTACAGACGAAGAGGAGATCTGGAC (SEQ ID NO: 266 ofUS20160369298; herein SEQ ID NO: 1724),TGGGACAATGGCGGTCGTCTCTCAGAGTTKTKKT (SEQ ID NO: 267 of US20160369298;herein SEQ ID NO: 1725), AGAGGACCKKTCCTCGATGGTTCATGGTGGAGTTA (SEQ ID NO:268 of US20160369298; herein SEQ ID NO: 1726),CCACTTAGGGCCTGGTCGATACCGTTCGGTG (SEQ ID NO: 269 of US20160369298; hereinSEQ ID NO: 1727), and TCTCGCCCCAAGAGTAGAAACCCTTCSTTYYG (SEQ ID NO: 270of US20160369298; herein SEQ ID NO: 1728).

In some embodiments, the AAV serotype may comprise an ocular celltargeting peptide as described in International Patent PublicationWO2016134375, the contents of which are herein incorporated by referencein their entirety, such as, but not limited to SEQ ID NO: 9, and SEQ IDNO:10 of WO2016134375. Further, any of the ocular cell targetingpeptides or amino acids described in WO2016134375, may be inserted intoany parent AAV serotype, such as, but not limited to, AAV2 (SEQ ID NO:8of WO2016134375; herein SEQ ID NO: 1729), or AAV9 (SEQ ID NO: 11 ofWO2016134375; herein SEQ ID NO: 1730). In some embodiments,modifications, such as insertions are made in AAV2 proteins at P34-A35,T138-A139, A139-P140, G453-T454, N587-R588, and/or R588-Q589. In certainembodiments, insertions are made at D384, G385, 1560, T561, N562, E563,E564, E565, N704, and/or Y705 of AAV9. The ocular cell targeting peptidemay be, but is not limited to, any of the following amino acidsequences, GSTPPPM (SEQ ID NO: 1 of WO2016134375; herein SEQ ID NO:1731), or GETRAPL (SEQ ID NO: 4 of WO2016134375; herein SEQ ID NO:1732).

In some embodiments, the AAV serotype may be modified as described inthe United States Publication US 20170145405 the contents of which areherein incorporated by reference in their entirety. AAV serotypes mayinclude, modified AAV2 (e.g., modifications at Y444F, Y500F, Y730Fand/or S662V), modified AAV3 (e.g., modifications at Y705F, Y731F and/orT492V), and modified AAV6 (e.g., modifications at S663V and/or T492V).

In some embodiments, the AAV serotype may be modified as described inthe International Publication WO2017083722 the contents of which areherein incorporated by reference in their entirety. AAV serotypes mayinclude, AAV1 (Y705+731F+T492V), AAV2 (Y444+500+730F+T491V), AAV3(Y705+731F), AAV5, AAV 5(Y436+693+719F), AAV6 (VP3 variantY705F/Y731F/T492V), AAV8 (Y733F), AAV9, AAV9 (VP3 variant Y731F), andAAV10 (Y733F).

In some embodiments, the AAV serotype may comprise, as described inInternational Patent Publication WO2017015102, the contents of which areherein incorporated by reference in their entirety, an engineeredepitope comprising the amino acids SPAKFA (SEQ ID NO: 24 ofWO2017015102; herein SEQ ID NO: 1733) or NKDKLN (SEQ ID NO:2 ofWO2017015102; herein SEQ ID NO: 1734). The epitope may be inserted inthe region of amino acids 665 to 670 based on the numbering of the VP1capsid of AAV8 (SEQ ID NO:3 of WO2017015102) and/or residues 664 to 668of AAV3B (SEQ ID NO:3).

In one embodiment, the AAV serotype may be, or may have a sequence asdescribed in International Patent Publication WO2017058892, the contentsof which are herein incorporated by reference in their entirety, suchas, but not limited to, AAV variants with capsid proteins that maycomprise a substitution at one or more (e.g., 2, 3, 4, 5, 6, or 7) ofamino acid residues 262-268, 370-379, 451-459, 472-473, 493-500,528-534, 547-552, 588-597, 709-710, 716-722 of AAV1, in any combination,or the equivalent amino acid residues in AAV2, AAV3, AAV4, AAV5, AAV6,AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAVrh8, AAVrh10, AAVrh32.33,bovine AAV or avian AAV. The amino acid substitution may be, but is notlimited to, any of the amino acid sequences described in WO2017058892.In one embodiment, the AAV may comprise an amino acid substitution atresidues 256L, 258K, 259Q, 261S, 263A, 264S, 265T, 266G, 272H, 385S,386Q, S472R, V473D, N500E 547S, 709A, 710N, 716D, 717N, 718N, 720L,A456T, Q457T, N458Q, K459S, T492S, K493A, S586R, S587G, S588N, T589Rand/or 722T of AAV1 (SEQ ID NO: 1 of WO2017058892) in any combination,244N, 246Q, 248R, 249E, 2501, 251K, 252S, 253G, 254S, 255V, 256D, 263Y,377E, 378N, 453L, 456R, 532Q, 533P, 535N, 536P, 537G, 538T, 539T, 540A,541T, 542Y, 543L, 546N, 653V, 654P, 656S, 697Q, 698F, 704D, 705S, 706T,707G, 708E, 709Y and/or 710R of AAV5 (SEQ ID NO:5 of WO2017058892) inany combination, 248R, 316V, 317Q, 318D, 319S, 443N, 530N, 5315, 532Q533P, 534A, 535N, 540A, 541 T, 542Y, 543L, 545G, 546N, 697Q, 704D, 706T,708E, 709Y and/or 710R of AAV5 (SEQ ID NO: 5 of WO2017058892) in anycombination, 264S, 266G, 269N, 272H, 457Q, 588S and/or 5891 of AAV6 (SEQID NO:6 WO2017058892) in any combination, 457T, 459N, 496G, 499N, SOON,589Q, 590N and/or 592A of AAV8 (SEQ ID NO: 8 WO2017058892) in anycombination, 451I, 452N, 453G, 454S, 455G, 456Q, 457N and/or 458Q ofAAV9 (SEQ ID NO: 9 WO2017058892) in any combination.

In some embodiments, the AAV may include a sequence of amino acids atpositions 155, 156 and 157 of VP1 or at positions 17, 18, 19 and 20 ofVP2, as described in International Publication No. WO 2017066764, thecontents of which are herein incorporated by reference in theirentirety. The sequences of amino acid may be, but not limited to, N-S-S,S-X-S, S-S-Y, N-X-S, N-S-Y, S-X-Y and N-X-Y, where N, X and Y are, butnot limited to, independently non-serine, or non-threonine amino acids,wherein the AAV may be, but not limited to AAV1, AAV2, AAV3, AAV4, AAV5,AAV6, AAV7, AAV8, AAV9, AAV10, AAV11 and AAV12. In some embodiments, theAAV may include a deletion of at least one amino acid at positions 156,157 or 158 of VP1 or at positions 19, 20 or 21 of VP2, wherein the AAVmay be, but not limited to AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7,AAV8, AAV9, AAV10, AAV11 and AAV12.

Viral Genome Component: Inverted Terminal Repeats (ITRs)

The AAV particles of the present disclosure comprise a viral genome withat least one ITR region and a payload region. In one embodiment, theviral genome has two ITRs. These two ITRs flank the payload region atthe 5′ and 3′ ends. The ITRs function as origins of replicationcomprising recognition sites for replication. ITRs comprise sequenceregions which can be complementary and symmetrically arranged. ITRsincorporated into viral genomes of the disclosure may be comprised ofnaturally occurring polynucleotide sequences or recombinantly derivedpolynucleotide sequences.

The ITRs may be derived from the same serotype as the capsid, selectedfrom any of the serotypes listed in Table 1, or a derivative thereof.The ITR may be of a different serotype than the capsid. In oneembodiment, the AAV particle has more than one ITR. In a non-limitingexample, the AAV particle has a viral genome comprising two ITRs. In oneembodiment, the ITRs are of the same serotype as one another. In anotherembodiment, the ITRs are of different serotypes. Non-limiting examplesinclude zero, one or both of the ITRs having the same serotype as thecapsid. In one embodiment both ITRs of the viral genome of the AAVparticle are AAV2 ITRs.

Independently, each ITR may be about 100 to about 150 nucleotides inlength. An ITR may be about 100-105 nucleotides in length, 106-110nucleotides in length, 111-115 nucleotides in length, 116-120nucleotides in length, 121-125 nucleotides in length, 126-130nucleotides in length, 131-135 nucleotides in length, 136-140nucleotides in length, 141-145 nucleotides in length or 146-150nucleotides in length. In one embodiment, the ITRs are 140-142nucleotides in length. Non-limiting examples of ITR length are 102, 140,141, 142, 145 nucleotides in length, and those having at least 95%identity thereto.

Viral Genome Component: Promoters

In one embodiment, the payload region of the viral genome comprises atleast one element to enhance the transgene target specificity andexpression (See e.g., Powell et al. Viral Expression Cassette Elementsto Enhance Transgene Target Specificity and Expression in Gene Therapy,2015; the contents of which are herein incorporated by reference in itsentirety). Non-limiting examples of elements to enhance the transgenetarget specificity and expression include promoters, endogenous miRNAs,post-transcriptional regulatory elements (PREs), polyadenylation (PolyA)signal sequences and upstream enhancers (USEs), CMV enhancers andintrons.

A person skilled in the art may recognize that expression of thepolypeptides of the disclosure in a target cell may require a specificpromoter, including but not limited to, a promoter that is speciesspecific, inducible, tissue-specific, or cell cycle-specific (Parr etal., Nat. Med. 3:1145-9 (1997); the contents of which are hereinincorporated by reference in their entirety).

In one embodiment, the promoter is deemed to be efficient when it drivesexpression of the polypeptide(s) encoded in the payload region of theviral genome of the AAV particle. As a non-limiting example, thatpolypeptide is AADC.

In one embodiment, the promoter is a promoter deemed to be efficientwhen it drives expression in the cell being targeted.

In one embodiment, the promoter is a promoter having a tropism for thecell being targeted.

In one embodiment, the promoter drives expression of the payload for aperiod of time in targeted tissues. Expression driven by a promoter maybe for a period of 1 hour, 2, hours, 3 hours, 4 hours, 5 hours, 6 hours,7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21hours, 22 hours, 23 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6days, 1 week, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 2weeks, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 3 weeks, 22days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30days, 31 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 13months, 14 months, 15 months, 16 months, 17 months, 18 months, 19months, 20 months, 21 months, 22 months, 23 months, 2 years, 3 years, 4years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years or morethan 10 years. Expression may be for 1-5 hours, 1-12 hours, 1-2 days,1-5 days, 1-2 weeks, 1-3 weeks, 1-4 weeks, 1-2 months, 1-4 months, 1-6months, 2-6 months, 3-6 months, 3-9 months, 4-8 months, 6-12 months, 1-2years, 1-5 years, 2-5 years, 3-6 years, 3-8 years, 4-8 years or 5-10years. As a non-limiting example, the promoter is a weak promoter forsustained expression of a payload in nervous tissues.

In one embodiment, the promoter drives expression of the polypeptides ofthe disclosure for at least 1 month, 2 months, 3 months, 4 months, 5months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1year, 2 years, 3 years 4 years, 5 years, 6 years, 7 years, 8 years, 9years, 10 years, 11 years, 12 years, 13 years, 14 years, 15 years, 16years, 17 years, 18 years, 19 years, 20 years, 21 years, 22 years, 23years, 24 years, 25 years, 26 years, 27 years, 28 years, 29 years, 30years, 31 years, 32 years, 33 years, 34 years, 35 years, 36 years, 37years, 38 years, 39 years, 40 years, 41 years, 42 years, 43 years, 44years, 45 years, 46 years, 47 years, 48 years, 49 years, 50 years, 55years, 60 years, 65 years, or more than 65 years.

Promoters may be naturally occurring or non-naturally occurring.Non-limiting examples of promoters include viral promoters, plantpromoters and mammalian promoters. In some embodiments, the promotersmay be human promoters. In some embodiments, the promoter may betruncated.

Promoters which drive or promote expression in most tissues include, butare not limited to, human elongation factor 1a-subunit (EF1α),cytomegalovirus (CMV) immediate-early enhancer and/or promoter, chickenβ-actin (CBA) and its derivative CAG, β glucuronidase (GUSB), orubiquitin C (UBC). Tissue-specific expression elements can be used torestrict expression to certain cell types such as, but not limited to,muscle specific promoters, B cell promoters, monocyte promoters,leukocyte promoters, macrophage promoters, pancreatic acinar cellpromoters, endothelial cell promoters, lung tissue promoters, astrocytepromoters, or nervous system promoters which can be used to restrictexpression to neurons, astrocytes, or oligodendrocytes.

Non-limiting examples of muscle-specific promoters include mammalianmuscle creatine kinase (MCK) promoter, mammalian desmin (DES) promoter,mammalian troponin I (TNNI2) promoter, and mammalian skeletalalpha-actin (ASKA) promoter (see, e.g. U.S. Patent Publication US20110212529, the contents of which are herein incorporated by referencein their entirety).

Non-limiting examples of tissue-specific expression elements for neuronsinclude neuron-specific enolase (NSE), platelet-derived growth factor(PDGF), platelet-derived growth factor B-chain (PDGF-β), synapsin (Syn),methyl-CpG binding protein 2 (MeCP2), Ca²⁺/calmodulin-dependent proteinkinase II (CaMKII), metabotropic glutamate receptor 2 (mGluR2),neurofilament light (NFL) or heavy (NFH), β-globin minigene nβ2,preproenkephalin (PPE), enkephalin (Enk) and excitatory amino acidtransporter 2 (EAAT2) promoters. Non-limiting examples oftissue-specific expression elements for astrocytes include glialfibrillary acidic protein (GFAP) and EAAT2 promoters. A non-limitingexample of a tissue-specific expression element for oligodendrocytesincludes the myelin basic protein (MBP) promoter.

In one embodiment, the promoter may be less than 1 kb. The promoter mayhave a length of 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300,310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440,450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580,590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720,730, 740, 750, 760, 770, 780, 790, 800 or more than 800 nucleotides. Thepromoter may have a length between 200-300, 200-400, 200-500, 200-600,200-700, 200-800, 300-400, 300-500, 300-600, 300-700, 300-800, 400-500,400-600, 400-700, 400-800, 500-600, 500-700, 500-800, 600-700, 600-800or 700-800.

In one embodiment, the promoter may be a combination of two or morecomponents of the same or different starting or parental promoters suchas, but not limited to, CMV and CBA. Each component may have a length of200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330,340, 350, 360, 370, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389,390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520,530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660,670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800 ormore than 800. Each component may have a length between 200-300,200-400, 200-500, 200-600, 200-700, 200-800, 300-400, 300-500, 300-600,300-700, 300-800, 400-500, 400-600, 400-700, 400-800, 500-600, 500-700,500-800, 600-700, 600-800 or 700-800. In one embodiment, the promoter isa combination of a 382 nucleotide CMV-enhancer sequence and a 260nucleotide CBA-promoter sequence.

In one embodiment, the viral genome comprises a ubiquitous promoter.Non-limiting examples of ubiquitous promoters include CMV, CBA(including derivatives CAG, CBh, etc.), EF-1α, PGK, UBC, GUSB (hGBp),and UCOE (promoter of HNRPA2B1-CBX3).

Yu et al. (Molecular Pain 2011, 7:63; the contents of which are hereinincorporated by reference in their entirety) evaluated the expression ofeGFP under the CAG, EFIα, PGK and UBC promoters in rat DRG cells andprimary DRG cells using lentiviral vectors and found that UBC showedweaker expression than the other 3 promoters and only 10-12% glialexpression was seen for all promoters. Soderblom et al. (E. Neuro 2015;the contents of which are herein incorporated by reference in itsentirety) evaluated the expression of eGFP in AAV8 with CMV and UBCpromoters and AAV2 with the CMV promoter after injection in the motorcortex. Intranasal administration of a plasmid containing a UBC or EFIαpromoter showed a sustained airway expression greater than theexpression with the CMV promoter (See e.g., Gill et al., Gene Therapy2001, Vol. 8, 1539-1546; the contents of which are herein incorporatedby reference in their entirety). Husain et al. (Gene Therapy 2009; thecontents of which are herein incorporated by reference in its entirety)evaluated an HβH construct with a hGUSB promoter, a HSV-1LAT promoterand an NSE promoter and found that the HβH construct showed weakerexpression than NSE in mouse brain. Passini and Wolfe (J. Virol. 2001,12382-12392, the contents of which are herein incorporated by referencein its entirety) evaluated the long term effects of the HβH vectorfollowing an intraventricular injection in neonatal mice and found thatthere was sustained expression for at least 1 year. Low expression inall brain regions was found by Xu et al. (Gene Therapy 2001, 8,1323-1332; the contents of which are herein incorporated by reference intheir entirety) when NFL and NFH promoters were used as compared to theCMV-lacZ, CMV-luc, EF, GFAP, hENK, nAChR, PPE, PPE+wpre, NSE (0.3 kb),NSE (1.8 kb) and NSE (1.8 kb+wpre). Xu et al. found that the promoteractivity in descending order was NSE (1.8 kb), EF, NSE (0.3 kb), GFAP,CMV, hENK, PPE, NFL and NFH. NFL is a 650 nucleotide promoter and NFH isa 920 nucleotide promoter which are both absent in the liver but NFH isabundant in the sensory proprioceptive neurons, brain and spinal cordand NFH is present in the heart. SCN8A is a 470 nucleotide promoterwhich expresses throughout the DRG, spinal cord and brain withparticularly high expression seen in the hippocampal neurons andcerebellar Purkinje cells, cortex, thalamus and hypothalamus (See e.g.,Drews et al. Identification of evolutionary conserved, functionalnoncoding elements in the promoter region of the sodium channel geneSCN8A, Mamm Genome (2007) 18:723-731; and Raymond et al. Expression ofAlternatively Spliced Sodium Channel a-subunit genes, Journal ofBiological Chemistry (2004) 279(44) 46234-46241; the contents of each ofwhich are herein incorporated by reference in their entireties).

Any of promoters taught by the aforementioned Yu, Soderblom, Gill,Husain, Passini, Xu, Drews or Raymond may be used in the presentdisclosures.

In one embodiment, the promoter is not cell specific.

In one embodiment, the promoter is an ubiquitin c (UBC) promoter. TheUBC promoter may have a size of 300-350 nucleotides. As a non-limitingexample, the UBC promoter is 332 nucleotides.

In one embodiment, the promoter is a β-glucuronidase (GUSB) promoter.The GUSB promoter may have a size of 350-400 nucleotides. As anon-limiting example, the GUSB promoter is 378 nucleotides.

In one embodiment, the promoter is a neurofilament light (NFL) promoter.The NFL promoter may have a size of 600-700 nucleotides. As anon-limiting example, the NFL promoter is 650 nucleotides.

In one embodiment, the promoter is a neurofilament heavy (NFH) promoter.The NFH promoter may have a size of 900-950 nucleotides. As anon-limiting example, the NFH promoter is 920 nucleotides.

In one embodiment, the promoter is a SCN8A promoter. The SCN8A promotermay have a size of 450-500 nucleotides. As a non-limiting example, theSCN8A promoter is 470 nucleotides.

In one embodiment, the promoter is a frataxin (FXN) promoter. The FXNpromoter may also be referred to as the FRDA promoter.

In one embodiment, the promoter is a phosphoglycerate kinase 1 (PGK)promoter.

In one embodiment, the promoter is a chicken β-actin (CBA) promoter.

In one embodiment, the promoter is a cytomegalovirus (CMV) promoter.

In one embodiment, the promoter is a H1 promoter.

In one embodiment, the promoter is an engineered promoter.

In one embodiment, the promoter is a liver or a skeletal musclepromoter. Non-limiting examples of liver promoters include humanα-1-antitrypsin (hAAT) and thyroxine binding globulin (TBG).Non-limiting examples of skeletal muscle promoters include Desmin, MCKor synthetic C5-12.

In one embodiment, the promoter is a RNA pol III promoter. As anon-limiting example, the RNA pol III promoter is U6. As a non-limitingexample, the RNA pol III promoter is H1.

In one embodiment, the viral genome comprises two promoters. As anon-limiting example, the promoters are an EF1α promoter and a CMVpromoter.

In one embodiment, the viral genome comprises an enhancer element, apromoter and/or a 5′UTR intron. The enhancer element, also referred toherein as an “enhancer,” may be, but is not limited to, a CMV enhancer,the promoter may be, but is not limited to, a CMV, CBA, UBC, GUSB, NSE,Synapsin, MeCP2, and GFAP promoter and the 5′UTR/intron may be, but isnot limited to, SV40, and CBA-MVM. As a non-limiting example, theenhancer, promoter and/or intron used in combination may be: (1) CMVenhancer, CMV promoter, SV40 5′UTR intron; (2) CMV enhancer, CBApromoter, SV 40 5′UTR intron; (3) CMV enhancer, CBA promoter, CBA-MVM5′UTR intron; (4) UBC promoter; (5) GUSB promoter; (6) NSE promoter; (7)Synapsin promoter; (8) MeCP2 promoter and (9) GFAP promoter.

In one embodiment, the viral genome comprises an engineered promoter.

In another embodiment, the viral genome comprises a promoter from anaturally expressed protein.

In one embodiment, a region located approximately ˜5 kb upstream of thefirst exon of the payload in order to allow for expression of thepayload with the promoter. (See e.g., Puspasari et al. Long RangeRegulation of Human FXN Gene Expression, PLOS ONE, 2011; the contents ofwhich is herein incorporated by reference in its entirety; a 17 bpregion located approximately 4.9 kb upstream of the first exon of thefrataxin gene in order to allow for expression with the FRDA promoter).

In one embodiment, the vector genome may comprise a promoter such as,but not limited to, CMV or U6. As a non-limiting example, the promoterfor the AAV particles comprising the payload of the present disclosureis a CMV promoter. As another non-limiting example, the promoter for theAAV particles comprising the payload of the present disclosure is a U6promoter.

In one embodiment, the vector genome may comprise a CMV and a U6promoter.

In one embodiment, the vector genome may comprise a CBA promoter.

Viral Genome Component: Untranslated Regions (UTRs)

By definition, wild type untranslated regions (UTRs) of a gene aretranscribed but not translated. Generally, the 5′ UTR starts at thetranscription start site and ends at the start codon and the 3′ UTRstarts immediately following the stop codon and continues until thetermination signal for transcription.

Features typically found in abundantly expressed genes of specifictarget organs may be engineered into UTRs to enhance the stability andprotein production. As a non-limiting example, a 5′ UTR from mRNAnormally expressed in the liver (e.g., albumin, serum amyloid A,Apolipoprotein AB/E, transferrin, alpha fetoprotein, erythropoietin, orFactor VIII) may be used in the viral genomes of the AAV particles ofthe disclosure to enhance expression in hepatic cell lines or liver.

While not wishing to be bound by theory, wild-type 5′ untranslatedregions (UTRs) include features which play roles in translationinitiation. Kozak sequences, which are commonly known to be involved inthe process by which the ribosome initiates translation of many genes,are usually included in 5′ UTRs. Kozak sequences have the consensusCCR(A/G)CCAUGG, where R is a purine (adenine or guanine) three basesupstream of the start codon (ATG), which is followed by another ‘G’.

In one embodiment, the 5′UTR in the viral genome includes a Kozaksequence.

In one embodiment, the 5′UTR in the viral genome does not include aKozak sequence.

While not wishing to be bound by theory, wild-type 3′ UTRs are known tohave stretches of Adenosines and Uridines embedded therein. These AUrich signatures are particularly prevalent in genes with high rates ofturnover. Based on their sequence features and functional properties,the AU rich elements (AREs) can be separated into three classes (Chen etal, 1995, the contents of which are herein incorporated by reference inits entirety): Class I AREs, such as, but not limited to, c-Myc andMyoD, contain several dispersed copies of an AUUUA motif within U-richregions. Class II AREs, such as, but not limited to, GM-CSF and TNF-α,possess two or more overlapping UUAUUUA(U/A)(U/A) nonamers. Class IIIARES, such as, but not limited to, c-Jun and Myogenin, are less welldefined. These U rich regions do not contain an AUUUA motif. Mostproteins binding to the AREs are known to destabilize the messenger,whereas members of the ELAV family, most notably HuR, have beendocumented to increase the stability of mRNA. HuR binds to AREs of allthe three classes. Engineering the HuR specific binding sites into the3′ UTR of nucleic acid molecules will lead to HuR binding and thus,stabilization of the message in vivo.

Introduction, removal or modification of 3′ UTR AU rich elements (AREs)can be used to modulate the stability of polynucleotides. Whenengineering specific polynucleotides, e.g., payload regions of viralgenomes, one or more copies of an ARE can be introduced to makepolynucleotides less stable and thereby curtail translation and decreaseproduction of the resultant protein. Likewise, AREs can be identifiedand removed or mutated to increase the intracellular stability and thusincrease translation and production of the resultant protein.

In one embodiment, the 3′ UTR of the viral genome may include anoligo(dT) sequence for templated addition of a poly-A tail.

In one embodiment, the viral genome may include at least one miRNA seed,binding site or full sequence. microRNAs (or miRNA or miR) are 19-25nucleotide noncoding RNAs that bind to the sites of nucleic acid targetsand down-regulate gene expression either by reducing nucleic acidmolecule stability or by inhibiting translation. A microRNA sequencecomprises a “seed” region, i.e., a sequence in the region of positions2-8 of the mature microRNA, which sequence has perfect Watson-Crickcomplementarity to the miRNA target sequence of the nucleic acid.

In one embodiment, the viral genome may be engineered to include, alteror remove at least one miRNA binding site, sequence or seed region.

Any UTR from any gene known in the art may be incorporated into theviral genome of the AAV particle. These UTRs, or portions thereof, maybe placed in the same orientation as in the gene from which they wereselected or they may be altered in orientation or location. In oneembodiment, the UTR used in the viral genome of the AAV particle may beinverted, shortened, lengthened, made with one or more other 5′ UTRs or3′ UTRs known in the art. As used herein, the term “altered” as itrelates to a UTR, means that the UTR has been changed in some way inrelation to a reference sequence. For example, a 3′ or 5′ UTR may bealtered relative to a wild type or native UTR by the change inorientation or location as taught above or may be altered by theinclusion of additional nucleotides, deletion of nucleotides, swappingor transposition of nucleotides.

In one embodiment, the viral genome of the AAV particle comprises atleast one artificial UTRs which is not a variant of a wild type UTR.

In one embodiment, the viral genome of the AAV particle comprises UTRswhich have been selected from a family of transcripts whose proteinsshare a common function, structure, feature or property.

Viral Genome Component: Polyadenylation Sequence

In one embodiment, the viral genome of the AAV particles of the presentdisclosure comprise at least one polyadenylation sequence. The viralgenome of the AAV particle may comprise a polyadenylation sequencebetween the 3′ end of the payload coding sequence and the 5′ end of the3′ITR.

In one embodiment, the polyadenylation sequence or “polyA sequence” mayrange from absent to about 500 nucleotides in length. Thepolyadenylation sequence may be, but is not limited to, 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42,43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60,61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111,112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125,126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139,140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153,154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167,168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181,182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195,196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209,210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223,224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237,238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251,252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265,266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279,280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293,294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307,308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321,322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335,336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349,350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363,364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377,378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391,392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405,406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419,420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433,434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447,448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461,462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475,476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489,490, 491, 492, 493, 494, 495, 496, 497, 498, 499, and 500 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 50-100 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 50-150 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 50-160 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 50-200 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 60-100 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 60-150 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 60-160 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 60-200 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 70-100 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 70-150 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 70-160 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 70-200 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 80-100 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 80-150 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 80-160 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 80-200 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 90-100 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 90-150 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 90-160 nucleotides inlength.

In one embodiment, the polyadenylation sequence is 90-200 nucleotides inlength.

Viral Genome Component: Introns

In one embodiment, the payload region comprises at least one element toenhance the expression such as one or more introns or portions thereof.Non-limiting examples of introns include, MVM (67-97 bps), F.IXtruncated intron 1 (300 bps), β-globin SD/immunoglobulin heavy chainsplice acceptor (250 bps), adenovirus splice donor/immunoglobin spliceacceptor (500 bps), SV40 late splice donor/splice acceptor (19S/16S)(180 bps) and hybrid adenovirus splice donor/IgG splice acceptor (230bps).

In one embodiment, the intron or intron portion may be 100-500nucleotides in length. The intron may have a length of 80, 90, 100, 110,120, 130, 140, 150, 160, 170, 171, 172, 173, 174, 175, 176, 177, 178,179, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300,310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440,450, 460, 470, 480, 490 or 500. The intron may have a length between80-100, 80-120, 80-140, 80-160, 80-180, 80-200, 80-250, 80-300, 80-350,80-400, 80-450, 80-500, 200-300, 200-400, 200-500, 300-400, 300-500, or400-500.

In one embodiment, the vector genome comprises at least one element toenhance the transgene target specificity and expression (See e.g.,Powell et al. Viral Expression Cassette Elements to Enhance TransgeneTarget Specificity and Expression in Gene Therapy, 2015; the contents ofwhich are herein incorporated by reference in its entirety) such as anintron. Non-limiting examples of introns include, MVM (67-97 bps), F.IXtruncated intron 1 (300 bps), β-globin SD/immunoglobulin heavy chainsplice acceptor (250 bps), adenovirus splice donor/immunoglobin spliceacceptor (500 bps), SV40 late splice donor/splice acceptor (19S/16S)(180 bps) and hybrid adenovirus splice donor/IgG splice acceptor (230bps).

In one embodiment, the intron may be 100-500 nucleotides in length. Theintron may have a length of 80, 90, 100, 110, 120, 130, 140, 150, 160,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 190, 200, 210,220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350,360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490 or500. The intron may have a length between 80-100, 80-120, 80-140,80-160, 80-180, 80-200, 80-250, 80-300, 80-350, 80-400, 80-450, 80-500,200-300, 200-400, 200-500, 300-400, 300-500, or 400-500.

Viral Genome Component: Filler Sequence

In one embodiment, the viral genome comprises one or more fillersequences.

In one embodiment, the viral genome comprises one or more fillersequences in order to have the length of the viral genome be the optimalsize for packaging. As a non-limiting example, the viral genomecomprises at least one filler sequence in order to have the length ofthe viral genome be about 2.3 kb. As a non-limiting example, the viralgenome comprises at least one filler sequence in order to have thelength of the viral genome be about 4.6 kb.

In one embodiment, the viral genome is a single stranded (ss) viralgenome and comprises one or more filler sequences which have a lengthabout between 0.1 kb-3.8 kb, such as, but not limited to, 0.1 kb, 0.2kb, 0.3 kb, 0.4 kb, 0.5 kb, 0.6 kb, 0.7 kb, 0.8 kb, 0.9 kb, 1 kb, 1.1kb, 1.2 kb, 1.3 kb, 1.4 kb, 1.5 kb, 1.6 kb, 1.7 kb, 1.8 kb, 1.9 kb, 2kb, 2.1 kb, 2.2 kb, 2.3 kb, 2.4 kb, 2.5 kb, 2.6 kb, 2.7 kb, 2.8 kb, 2.9kb, 3 kb, 3.1 kb, 3.2 kb, 3.3 kb, 3.4 kb, 3.5 kb, 3.6 kb, 3.7 kb, or 3.8kb. As a non-limiting example, the total length filler sequence in thevector genome is 3.1 kb. As a non-limiting example, the total lengthfiller sequence in the vector genome is 2.7 kb. As a non-limitingexample, the total length filler sequence in the vector genome is 0.8kb. As a non-limiting example, the total length filler sequence in thevector genome is 0.4 kb. As a non-limiting example, the length of eachfiller sequence in the vector genome is 0.8 kb. As a non-limitingexample, the length of each filler sequence in the vector genome is 0.4kb.

In one embodiment, the viral genome is a self-complementary (sc) viralgenome and comprises one or more filler sequences which have a lengthabout between 0.1 kb-1.5 kb, such as, but not limited to, 0.1 kb, 0.2kb, 0.3 kb, 0.4 kb, 0.5 kb, 0.6 kb, 0.7 kb, 0.8 kb, 0.9 kb, 1 kb, 1.1kb, 1.2 kb, 1.3 kb, 1.4 kb, or 1.5 kb. As a non-limiting example, thetotal length filler sequence in the vector genome is 0.8 kb. As anon-limiting example, the total length filler sequence in the vectorgenome is 0.4 kb. As a non-limiting example, the length of each fillersequence in the vector genome is 0.8 kb. As a non-limiting example, thelength of each filler sequence in the vector genome is 0.4 kb

In one embodiment, the viral genome comprises any portion of a fillersequence. The viral genome may comprise 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%,9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,75%, 80%, 85%, 90%, 95%, or 99% of a filler sequence.

In one embodiment, the viral genome is a single stranded (ss) viralgenome and comprises one or more filler sequences in order to have thelength of the viral genome be about 4.6 kb. As a non-limiting example,the viral genome comprises at least one filler sequence and the fillersequence is located 3′ to the 5′ ITR sequence. As a non-limitingexample, the viral genome comprises at least one filler sequence and thefiller sequence is located 5′ to a promoter sequence. As a non-limitingexample, the viral genome comprises at least one filler sequence and thefiller sequence is located 3′ to the polyadenylation signal sequence. Asa non-limiting example, the viral genome comprises at least one fillersequence and the filler sequence is located 5′ to the 3′ ITR sequence.As a non-limiting example, the viral genome comprises at least onefiller sequence, and the filler sequence is located between two intronsequences. As a non-limiting example, the viral genome comprises atleast one filler sequence, and the filler sequence is located within anintron sequence. As a non-limiting example, the viral genome comprisestwo filler sequences, and the first filler sequence is located 3′ to the5′ ITR sequence and the second filler sequence is located 3′ to thepolyadenylation signal sequence. As a non-limiting example, the viralgenome comprises two filler sequences, and the first filler sequence islocated 5′ to a promoter sequence and the second filler sequence islocated 3′ to the polyadenylation signal sequence. As a non-limitingexample, the viral genome comprises two filler sequences, and the firstfiller sequence is located 3′ to the 5′ ITR sequence and the secondfiller sequence is located 5′ to the 5′ ITR sequence.

In one embodiment, the viral genome is a self-complementary (sc) viralgenome and comprises one or more filler sequences in order to have thelength of the viral genome be about 2.3 kb. As a non-limiting example,the viral genome comprises at least one filler sequence and the fillersequence is located 3′ to the 5′ ITR sequence. As a non-limitingexample, the viral genome comprises at least one filler sequence and thefiller sequence is located 5′ to a promoter sequence. As a non-limitingexample, the viral genome comprises at least one filler sequence and thefiller sequence is located 3′ to the polyadenylation signal sequence. Asa non-limiting example, the viral genome comprises at least one fillersequence and the filler sequence is located 5′ to the 3′ ITR sequence.As a non-limiting example, the viral genome comprises at least onefiller sequence, and the filler sequence is located between two intronsequences. As a non-limiting example, the viral genome comprises atleast one filler sequence, and the filler sequence is located within anintron sequence. As a non-limiting example, the viral genome comprisestwo filler sequences, and the first filler sequence is located 3′ to the5′ ITR sequence and the second filler sequence is located 3′ to thepolyadenylation signal sequence. As a non-limiting example, the viralgenome comprises two filler sequences, and the first filler sequence islocated 5′ to a promoter sequence and the second filler sequence islocated 3′ to the polyadenylation signal sequence. As a non-limitingexample, the viral genome comprises two filler sequences, and the firstfiller sequence is located 3′ to the 5′ ITR sequence and the secondfiller sequence is located 5′ to the 5′ ITR sequence.

In one embodiment, the viral genome may comprise one or more fillersequences between one of more regions of the viral genome. In oneembodiment, the filler region may be located before a region such as,but not limited to, a payload region, an inverted terminal repeat (ITR),a promoter region, an intron region, an enhancer region, apolyadenylation signal sequence region, a multiple cloning site (MCS)region, and/or an exon region. In one embodiment, the filler region maybe located after a region such as, but not limited to, a payload region,an inverted terminal repeat (ITR), a promoter region, an intron region,an enhancer region, a polyadenylation signal sequence region, a multiplecloning site (MCS) region, and/or an exon region. In one embodiment, thefiller region may be located before and after a region such as, but notlimited to, a payload region, an inverted terminal repeat (ITR), apromoter region, an intron region, an enhancer region, a polyadenylationsignal sequence region, a multiple cloning site (MCS) region, and/or anexon region.

In one embodiment, the viral genome may comprise one or more fillersequences which bifurcates at least one region of the viral genome. Thebifurcated region of the viral genome may comprise 1%, 2%, 3%, 4%, 5%,6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of the of the region to the 5′of the filler sequence region. As a non-limiting example, the fillersequence may bifurcate at least one region so that 10% of the region islocated 5′ to the filler sequence and 90% of the region is located 3′ tothe filler sequence. As a non-limiting example, the filler sequence maybifurcate at least one region so that 20% of the region is located 5′ tothe filler sequence and 80% of the region is located 3′ to the fillersequence. As a non-limiting example, the filler sequence may bifurcateat least one region so that 30% of the region is located 5′ to thefiller sequence and 70% of the region is located 3′ to the fillersequence. As a non-limiting example, the filler sequence may bifurcateat least one region so that 40% of the region is located 5′ to thefiller sequence and 60% of the region is located 3′ to the fillersequence. As a non-limiting example, the filler sequence may bifurcateat least one region so that 50% of the region is located 5′ to thefiller sequence and 50% of the region is located 3′ to the fillersequence. As a non-limiting example, the filler sequence may bifurcateat least one region so that 60% of the region is located 5′ to thefiller sequence and 40% of the region is located 3′ to the fillersequence. As a non-limiting example, the filler sequence may bifurcateat least one region so that 70% of the region is located 5′ to thefiller sequence and 30% of the region is located 3′ to the fillersequence. As a non-limiting example, the filler sequence may bifurcateat least one region so that 80% of the region is located 5′ to thefiller sequence and 20% of the region is located 3′ to the fillersequence. As a non-limiting example, the filler sequence may bifurcateat least one region so that 90% of the region is located 5′ to thefiller sequence and 10% of the region is located 3′ to the fillersequence.

In one embodiment, the viral genome comprises a filler sequence afterthe 5′ ITR.

In one embodiment, the viral genome comprises a filler sequence afterthe promoter region. In one embodiment, the viral genome comprises afiller sequence after the payload region. In one embodiment, the viralgenome comprises a filler sequence after the intron region. In oneembodiment, the viral genome comprises a filler sequence after theenhancer region. In one embodiment, the viral genome comprises a fillersequence after the polyadenylation signal sequence region. In oneembodiment, the viral genome comprises a filler sequence after the MCSregion. In one embodiment, the viral genome comprises a filler sequenceafter the exon region.

In one embodiment, the viral genome comprises a filler sequence beforethe promoter region. In one embodiment, the viral genome comprises afiller sequence before the payload region. In one embodiment, the viralgenome comprises a filler sequence before the intron region. In oneembodiment, the viral genome comprises a filler sequence before theenhancer region. In one embodiment, the viral genome comprises a fillersequence before the polyadenylation signal sequence region. In oneembodiment, the viral genome comprises a filler sequence before the MCSregion. In one embodiment, the viral genome comprises a filler sequencebefore the exon region.

In one embodiment, the viral genome comprises a filler sequence beforethe 3′ ITR.

In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the 5′ ITR and the promoter region. In oneembodiment, a filler sequence may be located between two regions, suchas, but not limited to, the 5′ ITR and the payload region. In oneembodiment, a filler sequence may be located between two regions, suchas, but not limited to, the 5′ ITR and the intron region. In oneembodiment, a filler sequence may be located between two regions, suchas, but not limited to, the 5′ ITR and the enhancer region. In oneembodiment, a filler sequence may be located between two regions, suchas, but not limited to, the 5′ ITR and the polyadenylation signalsequence region. In one embodiment, a filler sequence may be locatedbetween two regions, such as, but not limited to, the 5′ ITR and the MCSregion.

In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the 5′ ITR and the exon region.

In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the promoter region and the payload region.In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the promoter region and the intron region.In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the promoter region and the enhancerregion. In one embodiment, a filler sequence may be located between tworegions, such as, but not limited to, the promoter region and thepolyadenylation signal sequence region. In one embodiment, a fillersequence may be located between two regions, such as, but not limitedto, the promoter region and the MCS region. In one embodiment, a fillersequence may be located between two regions, such as, but not limitedto, the promoter region and the exon region. In one embodiment, a fillersequence may be located between two regions, such as, but not limitedto, the promoter region and the 3′ ITR.

In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the payload region and the intron region.In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the payload region and the enhancer region.In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the payload region and the polyadenylationsignal sequence region. In one embodiment, a filler sequence may belocated between two regions, such as, but not limited to, the payloadregion and the MCS region. In one embodiment, a filler sequence may belocated between two regions, such as, but not limited to, the payloadregion and the exon region.

In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the payload region and the 3′ ITR.

In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the intron region and the enhancer region.In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the intron region and the polyadenylationsignal sequence region. In one embodiment, a filler sequence may belocated between two regions, such as, but not limited to, the intronregion and the MCS region. In one embodiment, a filler sequence may belocated between two regions, such as, but not limited to, the intronregion and the exon region. In one embodiment, a filler sequence may belocated between two regions, such as, but not limited to, the intronregion and the 3′ ITR. In one embodiment, a filler sequence may belocated between two regions, such as, but not limited to, the enhancerregion and the polyadenylation signal sequence region. In oneembodiment, a filler sequence may be located between two regions, suchas, but not limited to, the enhancer region and the MCS region. In oneembodiment, a filler sequence may be located between two regions, suchas, but not limited to, the enhancer region and the exon region. In oneembodiment, a filler sequence may be located between two regions, suchas, but not limited to, the enhancer region and the 3′ ITR.

In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the polyadenylation signal sequence regionand the MCS region. In one embodiment, a filler sequence may be locatedbetween two regions, such as, but not limited to, the polyadenylationsignal sequence region and the exon region. In one embodiment, a fillersequence may be located between two regions, such as, but not limitedto, the polyadenylation signal sequence region and the 3′ ITR.

In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the MCS region and the exon region. In oneembodiment, a filler sequence may be located between two regions, suchas, but not limited to, the MCS region and the 3′ ITR.

In one embodiment, a filler sequence may be located between two regions,such as, but not limited to, the exon region and the 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and promoterregion, and the second filler sequence may be located between thepromoter region and payload region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and promoter region, and the second fillersequence may be located between the promoter region and intron region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and promoterregion, and the second filler sequence may be located between thepromoter region and enhancer region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and promoter region, and the second fillersequence may be located between the promoter region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and promoter region, and the second filler sequence may belocated between the promoter region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and promoter region, and thesecond filler sequence may be located between the promoter region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand promoter region, and the second filler sequence may be locatedbetween the promoter region and 3′ ITR. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and promoter region, and the second fillersequence may be located between the payload region and intron region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and promoterregion, and the second filler sequence may be located between thepayload region and enhancer region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and promoter region, and the second fillersequence may be located between the payload region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and promoter region, and the second filler sequence may belocated between the payload region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and promoter region, and thesecond filler sequence may be located between the payload region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand promoter region, and the second filler sequence may be locatedbetween the payload region and 3′ ITR. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and promoter region, and the second fillersequence may be located between the intron region and enhancer region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and promoterregion, and the second filler sequence may be located between the intronregion and polyadenylation signal sequence region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and promoter region, and thesecond filler sequence may be located between the intron region and MCSregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand promoter region, and the second filler sequence may be locatedbetween the intron region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and promoter region, and the second fillersequence may be located between the intron region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and promoter region,and the second filler sequence may be located between the enhancerregion and polyadenylation signal sequence region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and promoter region, and thesecond filler sequence may be located between the enhancer region andMCS region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand promoter region, and the second filler sequence may be locatedbetween the enhancer region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and promoter region, and the second fillersequence may be located between the enhancer region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and promoter region,and the second filler sequence may be located between thepolyadenylation signal sequence region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and promoter region,and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and promoter region,and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and promoter region, and thesecond filler sequence may be located between the MCS region and exonregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand promoter region, and the second filler sequence may be locatedbetween the MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and promoter region, and the second filler sequencemay be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and payloadregion, and the second filler sequence may be located between thepromoter region and payload region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and payload region, and the second fillersequence may be located between the promoter region and intron region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and payloadregion, and the second filler sequence may be located between thepromoter region and enhancer region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and payload region, and the second fillersequence may be located between the promoter region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and payload region, and the second filler sequence may belocated between the promoter region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and payload region, and thesecond filler sequence may be located between the promoter region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand payload region, and the second filler sequence may be locatedbetween the promoter region and 3′ ITR. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and payload region, and the second fillersequence may be located between the payload region and intron region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and payloadregion, and the second filler sequence may be located between thepayload region and enhancer region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and payload region, and the second fillersequence may be located between the payload region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and payload region, and the second filler sequence may belocated between the payload region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and payload region, and thesecond filler sequence may be located between the payload region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand payload region, and the second filler sequence may be locatedbetween the payload region and 3′ ITR. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and payload region, and the second fillersequence may be located between the intron region and enhancer region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and payloadregion, and the second filler sequence may be located between the intronregion and polyadenylation signal sequence region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and payload region, and thesecond filler sequence may be located between the intron region and MCSregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand payload region, and the second filler sequence may be locatedbetween the intron region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and payload region, and the second fillersequence may be located between the intron region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and payload region,and the second filler sequence may be located between the enhancerregion and polyadenylation signal sequence region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and payload region, and thesecond filler sequence may be located between the enhancer region andMCS region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand payload region, and the second filler sequence may be locatedbetween the enhancer region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and payload region, and the second fillersequence may be located between the enhancer region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and payload region,and the second filler sequence may be located between thepolyadenylation signal sequence region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and payload region,and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and payload region,and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and payload region, and thesecond filler sequence may be located between the MCS region and exonregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand payload region, and the second filler sequence may be locatedbetween the MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and payload region, and the second filler sequencemay be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and intronregion, and the second filler sequence may be located between thepromoter region and payload region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and intron region, and the second fillersequence may be located between the promoter region and intron region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and intronregion, and the second filler sequence may be located between thepromoter region and enhancer region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and intron region, and the second fillersequence may be located between the promoter region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and intron region, and the second filler sequence may belocated between the promoter region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and intron region, and thesecond filler sequence may be located between the promoter region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand intron region, and the second filler sequence may be located betweenthe promoter region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and intron region, and the second filler sequence maybe located between the payload region and intron region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and intron region, andthe second filler sequence may be located between the payload region andenhancer region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between the5′ ITR and intron region, and the second filler sequence may be locatedbetween the payload region and polyadenylation signal sequence region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and intronregion, and the second filler sequence may be located between thepayload region and MCS region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and intron region, and the second filler sequence maybe located between the payload region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and intron region, andthe second filler sequence may be located between the payload region and3′ ITR. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand intron region, and the second filler sequence may be located betweenthe intron region and enhancer region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and intron region, and the second fillersequence may be located between the intron region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and intron region, and the second filler sequence may belocated between the intron region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and intron region, and thesecond filler sequence may be located between the intron region and exonregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand intron region, and the second filler sequence may be located betweenthe intron region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and intron region, and the second filler sequence maybe located between the enhancer region and polyadenylation signalsequence region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between the5′ ITR and intron region, and the second filler sequence may be locatedbetween the enhancer region and MCS region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and intron region, and the second fillersequence may be located between the enhancer region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and intronregion, and the second filler sequence may be located between theenhancer region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and intron region, and the second filler sequence maybe located between the polyadenylation signal sequence region and MCSregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand intron region, and the second filler sequence may be located betweenthe polyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and intron region, andthe second filler sequence may be located between the polyadenylationsignal sequence region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and intron region, and the second filler sequence maybe located between the MCS region and exon region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and intron region, and thesecond filler sequence may be located between the MCS region and 3′ ITR.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and intronregion, and the second filler sequence may be located between the exonregion and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and enhancerregion, and the second filler sequence may be located between thepromoter region and payload region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and enhancer region, and the second fillersequence may be located between the promoter region and intron region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and enhancerregion, and the second filler sequence may be located between thepromoter region and enhancer region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and enhancer region, and the second fillersequence may be located between the promoter region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and enhancer region, and the second filler sequence may belocated between the promoter region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and enhancer region, and thesecond filler sequence may be located between the promoter region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand enhancer region, and the second filler sequence may be locatedbetween the promoter region and 3′ ITR. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and enhancer region, and the second fillersequence may be located between the payload region and intron region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and enhancerregion, and the second filler sequence may be located between thepayload region and enhancer region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and enhancer region, and the second fillersequence may be located between the payload region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and enhancer region, and the second filler sequence may belocated between the payload region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and enhancer region, and thesecond filler sequence may be located between the payload region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand enhancer region, and the second filler sequence may be locatedbetween the payload region and 3′ ITR. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and enhancer region, and the second fillersequence may be located between the intron region and enhancer region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and enhancerregion, and the second filler sequence may be located between the intronregion and polyadenylation signal sequence region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and enhancer region, and thesecond filler sequence may be located between the intron region and MCSregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand enhancer region, and the second filler sequence may be locatedbetween the intron region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and enhancer region, and the second fillersequence may be located between the intron region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and enhancer region,and the second filler sequence may be located between the enhancerregion and polyadenylation signal sequence region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and enhancer region, and thesecond filler sequence may be located between the enhancer region andMCS region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand enhancer region, and the second filler sequence may be locatedbetween the enhancer region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and enhancer region, and the second fillersequence may be located between the enhancer region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and enhancer region,and the second filler sequence may be located between thepolyadenylation signal sequence region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and enhancer region,and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and enhancer region,and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and enhancer region, and thesecond filler sequence may be located between the MCS region and exonregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand enhancer region, and the second filler sequence may be locatedbetween the MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and enhancer region, and the second filler sequencemay be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR andpolyadenylation signal sequence region, and the second filler sequencemay be located between the promoter region and payload region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and polyadenylationsignal sequence region, and the second filler sequence may be locatedbetween the promoter region and intron region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and polyadenylation signalsequence region, and the second filler sequence may be located betweenthe promoter region and enhancer region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and polyadenylation signal sequenceregion, and the second filler sequence may be located between thepromoter region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and promoter regionand MCS region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between the5′ ITR and polyadenylation signal sequence region, and the second fillersequence may be located between the promoter region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR andpolyadenylation signal sequence region, and the second filler sequencemay be located between the promoter region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and polyadenylationsignal sequence region, and the second filler sequence may be locatedbetween the payload region and intron region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and polyadenylation signal sequenceregion, and the second filler sequence may be located between thepayload region and enhancer region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and polyadenylation signal sequence region,and the second filler sequence may be located between the payload regionand polyadenylation signal sequence region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and polyadenylation signal sequenceregion, and the second filler sequence may be located between thepayload region and MCS region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and polyadenylation signal sequence region, and thesecond filler sequence may be located between the payload region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand polyadenylation signal sequence region, and the second fillersequence may be located between the payload region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and polyadenylationsignal sequence region, and the second filler sequence may be locatedbetween the intron region and enhancer region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and polyadenylation signalsequence region, and the second filler sequence may be located betweenthe intron region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and intron region andMCS region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand polyadenylation signal sequence region, and the second fillersequence may be located between the intron region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR andpolyadenylation signal sequence region, and the second filler sequencemay be located between the intron region and 3′ ITR. In one embodiment,a viral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and polyadenylation signalsequence region, and the second filler sequence may be located betweenthe enhancer region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and polyadenylationsignal sequence region, and the second filler sequence may be locatedbetween the enhancer region and MCS region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and polyadenylation signal sequenceregion, and the second filler sequence may be located between theenhancer region and exon region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and polyadenylation signal sequence region, and thesecond filler sequence may be located between the enhancer region and 3′ITR. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand polyadenylation signal sequence region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and polyadenylation signal sequence region, and the secondfiller sequence may be located between the polyadenylation signalsequence region and exon region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and polyadenylation signal sequence region, and thesecond filler sequence may be located between the polyadenylation signalsequence region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and polyadenylation signal sequence region, and thesecond filler sequence may be located between the MCS region and exonregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand polyadenylation signal sequence region, and the second fillersequence may be located between the MCS region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and polyadenylationsignal sequence region, and the second filler sequence may be locatedbetween the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and MCS region,and the second filler sequence may be located between the promoterregion and payload region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and MCS region, and the second filler sequence may belocated between the promoter region and intron region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and MCS region, andthe second filler sequence may be located between the promoter regionand enhancer region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between the5′ ITR and MCS region, and the second filler sequence may be locatedbetween the promoter region and polyadenylation signal sequence region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and MCS region,and the second filler sequence may be located between the promoterregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and MCS region, and the second filler sequence may be locatedbetween the promoter region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and MCS region, and the second fillersequence may be located between the promoter region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and MCS region, andthe second filler sequence may be located between the payload region andintron region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand MCS region, and the second filler sequence may be located betweenthe payload region and enhancer region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and MCS region, and the second fillersequence may be located between the payload region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and MCS region, and the second filler sequence may be locatedbetween the payload region and MCS region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and MCS region, and the second fillersequence may be located between the payload region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and MCS region,and the second filler sequence may be located between the payload regionand 3′ ITR. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand MCS region, and the second filler sequence may be located betweenthe intron region and enhancer region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and MCS region, and the second fillersequence may be located between the intron region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and MCS region, and the second filler sequence may be locatedbetween the intron region and MCS region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and MCS region, and the second fillersequence may be located between the intron region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and MCS region,and the second filler sequence may be located between the intron regionand 3′ ITR. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand MCS region, and the second filler sequence may be located betweenthe enhancer region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and MCS region, andthe second filler sequence may be located between the enhancer regionand MCS region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between the5′ ITR and MCS region, and the second filler sequence may be locatedbetween the enhancer region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and MCS region, and the second fillersequence may be located between the enhancer region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and MCS region, andthe second filler sequence may be located between the polyadenylationsignal sequence region and MCS region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and MCS region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and MCS region, and the second filler sequence may be locatedbetween the polyadenylation signal sequence region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and MCS region, andthe second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand MCS region, and the second filler sequence may be located betweenthe MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and MCS region, and the second filler sequence may belocated between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and exon region,and the second filler sequence may be located between the promoterregion and payload region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and exon region, and the second filler sequence maybe located between the promoter region and intron region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and exon region, andthe second filler sequence may be located between the promoter regionand enhancer region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between the5′ ITR and exon region, and the second filler sequence may be locatedbetween the promoter region and polyadenylation signal sequence region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and exon region,and the second filler sequence may be located between the promoterregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe 5′ ITR and exon region, and the second filler sequence may belocated between the promoter region and exon region. In one embodiment,a viral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and exon region, and thesecond filler sequence may be located between the promoter region and 3′ITR. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand exon region, and the second filler sequence may be located betweenthe payload region and intron region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and exon region, and the second fillersequence may be located between the payload region and enhancer region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and exon region,and the second filler sequence may be located between the payload regionand polyadenylation signal sequence region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and exon region, and the second fillersequence may be located between the payload region and MCS region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and exon region,and the second filler sequence may be located between the payload regionand exon region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between the5′ ITR and exon region, and the second filler sequence may be locatedbetween the payload region and 3′ ITR. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the 5′ ITR and exon region, and the second fillersequence may be located between the intron region and enhancer region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and exon region,and the second filler sequence may be located between the intron regionand polyadenylation signal sequence region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and exon region, and the second fillersequence may be located between the intron region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and exon region, andthe second filler sequence may be located between the intron region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the 5′ ITRand exon region, and the second filler sequence may be located betweenthe intron region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and exon region, and the second filler sequence maybe located between the enhancer region and polyadenylation signalsequence region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between the5′ ITR and exon region, and the second filler sequence may be locatedbetween the enhancer region and MCS region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the 5′ ITR and exon region, and the second fillersequence may be located between the enhancer region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and exon region,and the second filler sequence may be located between the enhancerregion and 3′ ITR. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between the5′ ITR and exon region, and the second filler sequence may be locatedbetween the polyadenylation signal sequence region and MCS region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and exon region,and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the 5′ ITR and exon region, andthe second filler sequence may be located between the polyadenylationsignal sequence region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the 5′ ITR and exon region, and the second filler sequence maybe located between the MCS region and exon region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the 5′ ITR and exon region, and thesecond filler sequence may be located between the MCS region and 3′ ITR.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the 5′ ITR and exon region,and the second filler sequence may be located between the exon regionand 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andpayload region, and the second filler sequence may be located betweenthe payload region and intron region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the promoter region and payload region, and the secondfiller sequence may be located between the payload region and enhancerregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and payload region, and the second filler sequence may be locatedbetween the payload region and polyadenylation signal sequence region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andpayload region, and the second filler sequence may be located betweenthe payload region and MCS region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and payload region, and the second fillersequence may be located between the payload region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andpayload region, and the second filler sequence may be located betweenthe payload region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and payload region, and the second fillersequence may be located between the intron region and enhancer region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andpayload region, and the second filler sequence may be located betweenthe intron region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and payloadregion, and the second filler sequence may be located between the intronregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and payload region, and the second filler sequencemay be located between the intron region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and payloadregion, and the second filler sequence may be located between the intronregion and 3′ ITR. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between thepromoter region and payload region, and the second filler sequence maybe located between the enhancer region and polyadenylation signalsequence region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between thepromoter region and payload region, and the second filler sequence maybe located between the enhancer region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and payloadregion, and the second filler sequence may be located between theenhancer region and exon region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and payload region, and the second fillersequence may be located between the enhancer region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and payloadregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and payloadregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and payloadregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and payload region,and the second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and payload region, and the second filler sequence may be locatedbetween the MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and payload region, and the second fillersequence may be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andintron region, and the second filler sequence may be located between thepayload region and intron region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and intron region, and the second fillersequence may be located between the payload region and enhancer region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andintron region, and the second filler sequence may be located between thepayload region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and intronregion, and the second filler sequence may be located between thepayload region and MCS region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and intron region, and the second fillersequence may be located between the payload region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andintron region, and the second filler sequence may be located between thepayload region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and intron region, and the second fillersequence may be located between the intron region and enhancer region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andintron region, and the second filler sequence may be located between theintron region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and intronregion, and the second filler sequence may be located between the intronregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and intron region, and the second filler sequencemay be located between the intron region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and intronregion, and the second filler sequence may be located between the intronregion and 3′ ITR. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between thepromoter region and intron region, and the second filler sequence may belocated between the enhancer region and polyadenylation signal sequenceregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and intron region, and the second filler sequence may be locatedbetween the enhancer region and MCS region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the promoter region and intron region, and the secondfiller sequence may be located between the enhancer region and exonregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and intron region, and the second filler sequence may be locatedbetween the enhancer region and 3′ ITR. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the promoter region and intron region, and the secondfiller sequence may be located between the polyadenylation signalsequence region and MCS region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and intron region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and intron region, and the second filler sequencemay be located between the polyadenylation signal sequence region and 3′ITR. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and intron region, and the second filler sequence may be locatedbetween the MCS region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the promoter region and intron region, and the secondfiller sequence may be located between the MCS region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and intronregion, and the second filler sequence may be located between the exonregion and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andenhancer region, and the second filler sequence may be located betweenthe payload region and intron region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the promoter region and enhancer region, and the secondfiller sequence may be located between the payload region and enhancerregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and enhancer region, and the second filler sequence may belocated between the payload region and polyadenylation signal sequenceregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and enhancer region, and the second filler sequence may belocated between the payload region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and enhancer region,and the second filler sequence may be located between the payload regionand exon region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between thepromoter region and enhancer region, and the second filler sequence maybe located between the payload region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and enhancer region,and the second filler sequence may be located between the intron regionand enhancer region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between thepromoter region and enhancer region, and the second filler sequence maybe located between the intron region and polyadenylation signal sequenceregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and enhancer region, and the second filler sequence may belocated between the intron region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and enhancer region,and the second filler sequence may be located between the intron regionand exon region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between thepromoter region and enhancer region, and the second filler sequence maybe located between the intron region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and enhancer region,and the second filler sequence may be located between the enhancerregion and polyadenylation signal sequence region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and enhancer region,and the second filler sequence may be located between the enhancerregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and enhancer region, and the second filler sequencemay be located between the enhancer region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and enhancerregion, and the second filler sequence may be located between theenhancer region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and enhancer region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and enhancer region, and the second filler sequencemay be located between the polyadenylation signal sequence region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and enhancer region, and the second filler sequence may belocated between the polyadenylation signal sequence region and 3′ ITR.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andenhancer region, and the second filler sequence may be located betweenthe MCS region and exon region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and enhancer region, and the second fillersequence may be located between the MCS region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and enhancerregion, and the second filler sequence may be located between the exonregion and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the payload region and intron region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the payload region and enhancer region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the payload region and polyadenylation signalsequence region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between thepromoter region and polyadenylation signal sequence region, and thesecond filler sequence may be located between the payload region and MCSregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and polyadenylation signal sequence region, and the second fillersequence may be located between the payload region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the payload region and 3′ ITR. In one embodiment,a viral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and polyadenylationsignal sequence region, and the second filler sequence may be locatedbetween the intron region and enhancer region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and polyadenylationsignal sequence region, and the second filler sequence may be locatedbetween the intron region and polyadenylation signal sequence region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the intron region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the intron region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the intron region and 3′ ITR. In one embodiment,a viral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and polyadenylationsignal sequence region, and the second filler sequence may be locatedbetween the enhancer region and polyadenylation signal sequence region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the enhancer region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the enhancer region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the enhancer region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the polyadenylation signal sequence region andMCS region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and polyadenylation signal sequence region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and polyadenylation signal sequence region, and thesecond filler sequence may be located between the polyadenylation signalsequence region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and polyadenylation signal sequence region,and the second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and polyadenylation signal sequence region, and the second fillersequence may be located between the MCS region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andexon region, and the second filler sequence may be located between thepayload region and intron region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and exon region, and the second fillersequence may be located between the payload region and enhancer region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andexon region, and the second filler sequence may be located between thepayload region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and exonregion, and the second filler sequence may be located between thepayload region and MCS region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and exon region, and the second fillersequence may be located between the payload region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andexon region, and the second filler sequence may be located between thepayload region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and exon region, and the second fillersequence may be located between the intron region and enhancer region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andexon region, and the second filler sequence may be located between theintron region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and exonregion, and the second filler sequence may be located between the intronregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and exon region, and the second filler sequence maybe located between the intron region and exon region. In one embodiment,a viral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and exon region, andthe second filler sequence may be located between the intron region and3′ ITR. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and exon region, and the second filler sequence may be locatedbetween the enhancer region and polyadenylation signal sequence region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andexon region, and the second filler sequence may be located between theenhancer region and MCS region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and exon region, and the second fillersequence may be located between the enhancer region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region andexon region, and the second filler sequence may be located between theenhancer region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and exon region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and exon region, and the second filler sequence maybe located between the polyadenylation signal sequence region and exonregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and exon region, and the second filler sequence may be locatedbetween the polyadenylation signal sequence region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and exonregion, and the second filler sequence may be located between the MCSregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and exon region, and the second filler sequence maybe located between the MCS region and 3′ ITR. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the promoter region and exon region, and the secondfiller sequence may be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region and MCSregion, and the second filler sequence may be located between thepayload region and intron region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and MCS region, and the second fillersequence may be located between the payload region and enhancer region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region and MCSregion, and the second filler sequence may be located between thepayload region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and MCSregion, and the second filler sequence may be located between thepayload region and MCS region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and MCS region, and the second fillersequence may be located between the payload region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region and MCSregion, and the second filler sequence may be located between thepayload region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and MCS region, and the second fillersequence may be located between the intron region and enhancer region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region and MCSregion, and the second filler sequence may be located between the intronregion and polyadenylation signal sequence region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and MCS region, andthe second filler sequence may be located between the intron region andMCS region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and MCS region, and the second filler sequence may be locatedbetween the intron region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the promoter region and MCS region, and the secondfiller sequence may be located between the intron region and 3′ ITR. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region and MCSregion, and the second filler sequence may be located between theenhancer region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and MCSregion, and the second filler sequence may be located between theenhancer region and MCS region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and MCS region, and the second fillersequence may be located between the enhancer region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region and MCSregion, and the second filler sequence may be located between theenhancer region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and MCS region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and MCS region, and the second filler sequence maybe located between the polyadenylation signal sequence region and exonregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and MCS region, and the second filler sequence may be locatedbetween the polyadenylation signal sequence region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and MCSregion, and the second filler sequence may be located between the MCSregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and MCS region, and the second filler sequence maybe located between the MCS region and 3′ ITR. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the promoter region and MCS region, and the secondfiller sequence may be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region and3′ITR, and the second filler sequence may be located between the payloadregion and intron region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and 3′ITR, and the second filler sequence may belocated between the payload region and enhancer region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and 3′ITR,and the second filler sequence may be located between the payload regionand polyadenylation signal sequence region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the promoter region and 3′ITR, and the second fillersequence may be located between the payload region and MCS region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region and3′ITR, and the second filler sequence may be located between the payloadregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and 3′ITR, and the second filler sequence may belocated between the payload region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and 3′ITR, and thesecond filler sequence may be located between the intron region andenhancer region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between thepromoter region and 3′ITR, and the second filler sequence may be locatedbetween the intron region and polyadenylation signal sequence region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the promoter region and3′ITR, and the second filler sequence may be located between the intronregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and 3′ITR, and the second filler sequence may belocated between the intron region and exon region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and 3′ITR, and thesecond filler sequence may be located between the intron region and 3′ITR. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and 3′ITR, and the second filler sequence may be located betweenthe enhancer region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and 3′ITR,and the second filler sequence may be located between the enhancerregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe promoter region and 3′ITR, and the second filler sequence may belocated between the enhancer region and exon region. In one embodiment,a viral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and 3′ITR, and thesecond filler sequence may be located between the enhancer region and 3′ITR. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and 3′ITR, and the second filler sequence may be located betweenthe polyadenylation signal sequence region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and 3′ITR,and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the promoter region and 3′ITR,and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the promoter region and 3′ITR, and thesecond filler sequence may be located between the MCS region and exonregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the promoterregion and 3′ITR, and the second filler sequence may be located betweenthe MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the promoter region and 3′ITR, and the second filler sequencemay be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the payload region andintron region, and the second filler sequence may be located between theintron region and enhancer region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the payload region and intron region, and the second fillersequence may be located between the intron region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe payload region and intron region, and the second filler sequence maybe located between the intron region and MCS region. In one embodiment,a viral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and intron region,and the second filler sequence may be located between the intron regionand exon region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between thepayload region and intron region, and the second filler sequence may belocated between the intron region and 3′ ITR. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the payload region and intron region, and the secondfiller sequence may be located between the enhancer region andpolyadenylation signal sequence region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the payload region and intron region, and the secondfiller sequence may be located between the enhancer region and MCSregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and intron region, and the second filler sequence may be locatedbetween the enhancer region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the payload region and intron region, and the secondfiller sequence may be located between the enhancer region and 3′ ITR.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the payload region andintron region, and the second filler sequence may be located between thepolyadenylation signal sequence region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region and intronregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region and intronregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and intron region,and the second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and intron region, and the second filler sequence may be locatedbetween the MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the payload region and intron region, and the second fillersequence may be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the payload region andenhancer region, and the second filler sequence may be located betweenthe intron region and enhancer region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the payload region and enhancer region, and the secondfiller sequence may be located between the intron region andpolyadenylation signal sequence region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the payload region and enhancer region, and thesecond filler sequence may be located between the intron region and MCSregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and enhancer region, and the second filler sequence may belocated between the intron region and exon region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and enhancer region,and the second filler sequence may be located between the intron regionand 3′ ITR. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and enhancer region, and the second filler sequence may belocated between the enhancer region and polyadenylation signal sequenceregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and enhancer region, and the second filler sequence may belocated between the enhancer region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and enhancer region,and the second filler sequence may be located between the enhancerregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe payload region and enhancer region, and the second filler sequencemay be located between the enhancer region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region and enhancerregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region and enhancerregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region and enhancerregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and enhancer region,and the second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and enhancer region, and the second filler sequence may belocated between the MCS region and 3′ ITR. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the payload region and enhancer region, and thesecond filler sequence may be located between the exon region and 3′ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the payload region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the intron region and enhancer region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the intron region and polyadenylation signalsequence region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between thepayload region and polyadenylation signal sequence region, and thesecond filler sequence may be located between the intron region and MCSregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and polyadenylation signal sequence region, and the second fillersequence may be located between the intron region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the payload region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the intron region and 3′ ITR. In one embodiment,a viral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and polyadenylationsignal sequence region, and the second filler sequence may be locatedbetween the enhancer region and polyadenylation signal sequence region.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the payload region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the enhancer region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the enhancer region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the enhancer region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the polyadenylation signal sequence region andMCS region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and polyadenylation signal sequence region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe payload region and polyadenylation signal sequence region, and thesecond filler sequence may be located between the polyadenylation signalsequence region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the payload region and polyadenylation signal sequence region,and the second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and polyadenylation signal sequence region, and the second fillersequence may be located between the MCS region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the payload region and MCSregion, and the second filler sequence may be located between the intronregion and enhancer region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the payload region and MCS region, and the second fillersequence may be located between the intron region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe payload region and MCS region, and the second filler sequence may belocated between the intron region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and MCS region, andthe second filler sequence may be located between the intron region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and MCS region, and the second filler sequence may be locatedbetween the intron region and 3′ ITR. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the payload region and MCS region, and the second fillersequence may be located between the enhancer region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe payload region and MCS region, and the second filler sequence may belocated between the enhancer region and MCS region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and MCS region, andthe second filler sequence may be located between the enhancer regionand exon region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between thepayload region and MCS region, and the second filler sequence may belocated between the enhancer region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and MCS region, andthe second filler sequence may be located between the polyadenylationsignal sequence region and MCS region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the payload region and MCS region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe payload region and MCS region, and the second filler sequence may belocated between the polyadenylation signal sequence region and 3′ ITR.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the payload region and MCSregion, and the second filler sequence may be located between the MCSregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe payload region and MCS region, and the second filler sequence may belocated between the MCS region and 3′ ITR. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the payload region and MCS region, and the secondfiller sequence may be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the payload region and exonregion, and the second filler sequence may be located between the intronregion and enhancer region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the payload region and exon region, and the second fillersequence may be located between the intron region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe payload region and exon region, and the second filler sequence maybe located between the intron region and MCS region. In one embodiment,a viral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and exon region, andthe second filler sequence may be located between the intron region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and exon region, and the second filler sequence may be locatedbetween the intron region and 3′ ITR. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the payload region and exon region, and the secondfiller sequence may be located between the enhancer region andpolyadenylation signal sequence region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the payload region and exon region, and the secondfiller sequence may be located between the enhancer region and MCSregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and exon region, and the second filler sequence may be locatedbetween the enhancer region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the payload region and exon region, and the secondfiller sequence may be located between the enhancer region and 3′ ITR.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the payload region and exonregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region and exonregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region and exonregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and exon region, andthe second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and exon region, and the second filler sequence may be locatedbetween the MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the payload region and exon region, and the second fillersequence may be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the payload region and 3′ITR region, and the second filler sequence may be located between theintron region and enhancer region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the payload region and 3′ ITR region, and the second fillersequence may be located between the intron region and polyadenylationsignal sequence region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe payload region and 3′ ITR region, and the second filler sequence maybe located between the intron region and MCS region. In one embodiment,a viral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and 3′ ITR region,and the second filler sequence may be located between the intron regionand exon region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between thepayload region and 3′ ITR region, and the second filler sequence may belocated between the intron region and 3′ ITR. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the payload region and 3′ ITR region, and the secondfiller sequence may be located between the enhancer region andpolyadenylation signal sequence region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the payload region and 3′ ITR region, and the secondfiller sequence may be located between the enhancer region and MCSregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and 3′ ITR region, and the second filler sequence may be locatedbetween the enhancer region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the payload region and 3′ ITR region, and the secondfiller sequence may be located between the enhancer region and 3′ ITR.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the payload region and 3′ITR region, and the second filler sequence may be located between thepolyadenylation signal sequence region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region and 3′ ITRregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the payload region and 3′ ITRregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the payload region and 3′ ITR region,and the second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the payloadregion and 3′ ITR region, and the second filler sequence may be locatedbetween the MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the payload region and 3′ ITR region, and the second fillersequence may be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the intron region andenhancer region, and the second filler sequence may be located betweenthe enhancer region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the intron region and enhancerregion, and the second filler sequence may be located between theenhancer region and MCS region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the intron region and enhancer region, and the second fillersequence may be located between the enhancer region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the intron region andenhancer region, and the second filler sequence may be located betweenthe enhancer region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the intron region and enhancer region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe intron region and enhancer region, and the second filler sequencemay be located between the polyadenylation signal sequence region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the intronregion and enhancer region, and the second filler sequence may belocated between the polyadenylation signal sequence region and 3′ ITR.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the intron region andenhancer region, and the second filler sequence may be located betweenthe MCS region and exon region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the intron region and enhancer region, and the second fillersequence may be located between the MCS region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the intron region and enhancerregion, and the second filler sequence may be located between the exonregion and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the intron region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the enhancer region and polyadenylation signalsequence region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between theintron region and polyadenylation signal sequence region, and the secondfiller sequence may be located between the enhancer region and MCSregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the intronregion and polyadenylation signal sequence region, and the second fillersequence may be located between the enhancer region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the intron region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the enhancer region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the intron region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the polyadenylation signal sequence region andMCS region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the intronregion and polyadenylation signal sequence region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe intron region and polyadenylation signal sequence region, and thesecond filler sequence may be located between the polyadenylation signalsequence region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the intron region and polyadenylation signal sequence region,and the second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the intronregion and polyadenylation signal sequence region, and the second fillersequence may be located between the MCS region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the intron region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the intron region and MCSregion, and the second filler sequence may be located between theenhancer region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the intron region and MCS region,and the second filler sequence may be located between the enhancerregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe intron region and MCS region, and the second filler sequence may belocated between the enhancer region and exon region. In one embodiment,a viral genome may comprise two filler sequences, the first fillersequence may be located between the intron region and MCS region, andthe second filler sequence may be located between the enhancer regionand 3′ ITR. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the intronregion and MCS region, and the second filler sequence may be locatedbetween the polyadenylation signal sequence region and MCS region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the intron region and MCSregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the intron region and MCS region,and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the intron region and MCS region, andthe second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the intronregion and MCS region, and the second filler sequence may be locatedbetween the MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the intron region and MCS region, and the second filler sequencemay be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the intron region and exonregion, and the second filler sequence may be located between theenhancer region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the intron region and exonregion, and the second filler sequence may be located between theenhancer region and MCS region. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the intron region and exon region, and the second fillersequence may be located between the enhancer region and exon region. Inone embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the intron region and exonregion, and the second filler sequence may be located between theenhancer region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the intron region and exon region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and MCS region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe intron region and exon region, and the second filler sequence may belocated between the polyadenylation signal sequence region and exonregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the intronregion and exon region, and the second filler sequence may be locatedbetween the polyadenylation signal sequence region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the intron region and exonregion, and the second filler sequence may be located between the MCSregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe intron region and exon region, and the second filler sequence may belocated between the MCS region and 3′ ITR. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the intron region and exon region, and the secondfiller sequence may be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the intron region and3′ITR, and the second filler sequence may be located between theenhancer region and polyadenylation signal sequence region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the intron region and 3′ITR, andthe second filler sequence may be located between the enhancer regionand MCS region. In one embodiment, a viral genome may comprise twofiller sequences, the first filler sequence may be located between theintron region and 3′ITR, and the second filler sequence may be locatedbetween the enhancer region and exon region. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the intron region and 3′ITR, and the second fillersequence may be located between the enhancer region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the intron region and 3′ITR, andthe second filler sequence may be located between the polyadenylationsignal sequence region and MCS region. In one embodiment, a viral genomemay comprise two filler sequences, the first filler sequence may belocated between the intron region and 3′ITR, and the second fillersequence may be located between the polyadenylation signal sequenceregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe intron region and 3′ITR, and the second filler sequence may belocated between the polyadenylation signal sequence region and 3′ ITR.In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the intron region and3′ITR, and the second filler sequence may be located between the MCSregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe intron region and 3′ITR, and the second filler sequence may belocated between the MCS region and 3′ ITR. In one embodiment, a viralgenome may comprise two filler sequences, the first filler sequence maybe located between the intron region and 3′ITR, and the second fillersequence may be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the enhancer region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the polyadenylation signal sequence region andMCS region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the enhancerregion and polyadenylation signal sequence region, and the second fillersequence may be located between the polyadenylation signal sequenceregion and exon region. In one embodiment, a viral genome may comprisetwo filler sequences, the first filler sequence may be located betweenthe enhancer region and polyadenylation signal sequence region, and thesecond filler sequence may be located between the polyadenylation signalsequence region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the enhancer region and polyadenylation signal sequence region,and the second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the enhancerregion and polyadenylation signal sequence region, and the second fillersequence may be located between the MCS region and 3′ ITR. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the enhancer region andpolyadenylation signal sequence region, and the second filler sequencemay be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the enhancer region and MCSregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the enhancer region and MCSregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the enhancer region and MCSregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the enhancer region and MCS region, andthe second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the enhancerregion and MCS region, and the second filler sequence may be locatedbetween the MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the enhancer region and MCS region, and the second fillersequence may be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the enhancer region andexon region, and the second filler sequence may be located between thepolyadenylation signal sequence region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the enhancer region and exonregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the enhancer region and exonregion, and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the enhancer region and exon region, andthe second filler sequence may be located between the MCS region andexon region. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the enhancerregion and exon region, and the second filler sequence may be locatedbetween the MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the enhancer region and exon region, and the second fillersequence may be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the enhancer region and 3′ITR, and the second filler sequence may be located between thepolyadenylation signal sequence region and MCS region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the enhancer region and 3′ ITR,and the second filler sequence may be located between thepolyadenylation signal sequence region and exon region. In oneembodiment, a viral genome may comprise two filler sequences, the firstfiller sequence may be located between the enhancer region and 3′ ITR,and the second filler sequence may be located between thepolyadenylation signal sequence region and 3′ ITR. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the enhancer region and 3′ ITR, and thesecond filler sequence may be located between the MCS region and exonregion. In one embodiment, a viral genome may comprise two fillersequences, the first filler sequence may be located between the enhancerregion and 3′ ITR, and the second filler sequence may be located betweenthe MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the enhancer region and 3′ ITR, and the second filler sequencemay be located between the exon region and 3′ ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the polyadenylation signalsequence region and MCS region, and the second filler sequence may belocated between the MCS region and exon region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the polyadenylation signal sequenceregion and MCS region, and the second filler sequence may be locatedbetween the MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the polyadenylation signal sequence region and MCS region, andthe second filler sequence may be located between the exon region and 3′ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the polyadenylation signalsequence region and exon region, and the second filler sequence may belocated between the MCS region and exon region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the polyadenylation signal sequenceregion and exon region, and the second filler sequence may be locatedbetween the MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the polyadenylation signal sequence region and exon region, andthe second filler sequence may be located between the exon region and 3′ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the polyadenylation signalsequence region and 3′ ITR, and the second filler sequence may belocated between the MCS region and exon region. In one embodiment, aviral genome may comprise two filler sequences, the first fillersequence may be located between the polyadenylation signal sequenceregion and 3′ ITR, and the second filler sequence may be located betweenthe MCS region and 3′ ITR. In one embodiment, a viral genome maycomprise two filler sequences, the first filler sequence may be locatedbetween the polyadenylation signal sequence region and 3′ ITR, and thesecond filler sequence may be located between the exon region and 3′ITR.

In one embodiment, a viral genome may comprise two filler sequences, thefirst filler sequence may be located between the MCS region and exonregion, and the second filler sequence may be located between the exonregion and 3′ ITR.

AAV Production

The present disclosure provides methods for the generation of parvoviralparticles, e.g. AAV particles, by viral genome replication in a viralreplication cell.

In accordance with the disclosure, the viral genome comprising a payloadregion will be incorporated into the AAV particle produced in the viralreplication cell. Methods of making AAV particles are well known in theart and are described in e.g., U.S. Pat. Nos. 6,204,059, 5,756,283,6,258,595, 6,261,551, 6,270,996, 6,281,010, 6,365,394, 6,475,769,6,482,634, 6,485,966, 6,943,019, 6,953,690, 7,022,519, 7,238,526,7,291,498 and 7,491,508, 5,064,764, 6,194,191, 6,566,118, 8,137,948; orInternational Publication Nos. WO1996039530, WO1998010088, WO1999014354,WO1999015685, WO1999047691, WO2000055342, WO2000075353 and WO2001023597;Methods In Molecular Biology, ed. Richard, Humana Press, NJ (1995);O'Reilly et al., Baculovirus Expression Vectors, A Laboratory Manual,Oxford Univ. Press (1994); Samulski et al., J. Vir. 63:3822-8 (1989);Kajigaya et al., Proc. Nat'l. Acad. Sci. USA 88: 4646-50 (1991); Ruffinget al., J. Vir. 66:6922-30 (1992); Kimbauer et al., Vir., 219:37-44(1996); Zhao et al., Vir. 272:382-93 (2000); the contents of each ofwhich are herein incorporated by reference in their entirety. In oneembodiment, the AAV particles are made using the methods described inWO2015191508, the contents of which are herein incorporated by referencein their entirety.

Viral replication cells commonly used for production of recombinant AAVparticles include but are not limited to 293 cells, COS cells, HeLacells, KB cells, and other mammalian cell lines as described in U.S.Pat. Nos. 6,156,303, 5,387,484, 5,741,683, 5,691,176, and 5,688,676;U.S. patent publication No. 2002/0081721, and International PatentPublication Nos. WO 00/47757, WO 00/24916, and WO 96/17947, the contentsof each of which are herein incorporated by reference in theirentireties.

In some embodiments, the present disclosure provides a method forproducing an AAV particle having enhanced (increased, improved)transduction efficiency comprising the steps of: 1) co-transfectingcompetent bacterial cells with a bacmid vector and either a viralconstruct vector and/or AAV payload construct vector, 2) isolating theresultant viral construct expression vector and AAV payload constructexpression vector and separately transfecting viral replication cells,3) isolating and purifying resultant payload and viral constructparticles comprising viral construct expression vector or AAV payloadconstruct expression vector, 4) co-infecting a viral replication cellwith both the AAV payload and viral construct particles comprising viralconstruct expression vector or AAV payload construct expression vector,and 5) harvesting and purifying the AAV particle comprising a viralgenome.

In some embodiments, the present disclosure provides a method forproducing an AAV particle comprising the steps of 1) simultaneouslyco-transfecting mammalian cells, such as, but not limited to HEK293cells, with a payload region, a construct expressing rep and cap genesand a helper construct, 2) harvesting and purifying the AAV particlecomprising a viral genome.

In some embodiments, the viral genome of the AAV particle of thedisclosure optionally encodes a selectable marker. The selectable markermay comprise a cell-surface marker, such as any protein expressed on thesurface of the cell including, but not limited to receptors, CD markers,lectins, integrins, or truncated versions thereof.

In some embodiments, selectable marker reporter genes as described inInternational application No. WO 96/23810; Heim et al., Current Biology2:178-182 (1996); Heim et al., Proc. Natl. Acad. Sci. USA (1995); orHeim et al., Science 373:663-664 (1995); WO 96/30540, the contents ofeach of which are incorporated herein by reference in their entireties).

Genome Size

In one embodiment, the AAV particle which comprises a payload describedherein may be single stranded or double stranded vector genome. The sizeof the vector genome may be small, medium, large or the maximum size.Additionally, the vector genome may comprise a promoter and a polyAtail.

In one embodiment, the vector genome which comprises a payload describedherein may be a small single stranded vector genome. A small singlestranded vector genome may be 2.7 to 3.5 kb in size such as about 2.7,2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, and 3.5 kb in size. As a non-limitingexample, the small single stranded vector genome may be 3.2 kb in size.Additionally, the vector genome may comprise a promoter and a polyAtail.

In one embodiment, the vector genome which comprises a payload describedherein may be a small double stranded vector genome. A small doublestranded vector genome may be 1.3 to 1.7 kb in size such as about 1.3,1.4, 1.5, 1.6, and 1.7 kb in size. As a non-limiting example, the smalldouble stranded vector genome may be 1.6 kb in size. Additionally, thevector genome may comprise a promoter and a polyA tail.

In one embodiment, the vector genome which comprises a payload describedherein may be a medium single stranded vector genome. A medium singlestranded vector genome may be 3.6 to 4.3 kb in size such as about 3.6,3.7, 3.8, 3.9, 4.0, 4.1, 4.2 and 4.3 kb in size. As a non-limitingexample, the medium single stranded vector genome may be 4.0 kb in size.Additionally, the vector genome may comprise a promoter and a polyAtail.

In one embodiment, the vector genome which comprises a payload describedherein may be a medium double stranded vector genome. A medium doublestranded vector genome may be 1.8 to 2.1 kb in size such as about 1.8,1.9, 2.0, and 2.1 kb in size. As a non-limiting example, the mediumdouble stranded vector genome may be 2.0 kb in size. Additionally, thevector genome may comprise a promoter and a polyA tail.

In one embodiment, the vector genome which comprises a payload describedherein may be a large single stranded vector genome. A large singlestranded vector genome may be 4.4 to 6.0 kb in size such as about 4.4,4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8,5.9 and 6.0 kb in size. As a non-limiting example, the large singlestranded vector genome may be 4.7 kb in size. As another non-limitingexample, the large single stranded vector genome may be 4.8 kb in size.As yet another non-limiting example, the large single stranded vectorgenome may be 6.0 kb in size. Additionally, the vector genome maycomprise a promoter and a polyA tail.

In one embodiment, the vector genome which comprises a payload describedherein may be a large double stranded vector genome. A large doublestranded vector genome may be 2.2 to 3.0 kb in size such as about 2.2,2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9 and 3.0 kb in size. As a non-limitingexample, the large double stranded vector genome may be 2.4 kb in size.Additionally, the vector genome may comprise a promoter and a polyAtail.

Payloads

The AAV particles of the present disclosure comprise at least onepayload region. As used herein, “payload” or “payload region” refers toone or more polynucleotides or polynucleotide regions encoded by orwithin a viral genome or an expression product of such polynucleotide orpolynucleotide region, e.g., a transgene, a polynucleotide encoding apolypeptide or multi-polypeptide or a modulatory nucleic acid orregulatory nucleic acid. Payloads of the present disclosure typicallyencode polypeptides or fragments or variants thereof.

The payload region may be constructed in such a way as to reflect aregion similar to or mirroring the natural organization of an mRNA.

The payload region may comprise a combination of coding and non-codingnucleic acid sequences.

In some embodiments, the AAV payload region may encode a coding ornon-coding RNA.

In one embodiment, the AAV particle comprises a viral genome with apayload region comprising nucleic acid sequences encoding more than onepolypeptide of interest. In such an embodiment, a viral genome encodingmore than one polypeptide may be replicated and packaged into a viralparticle. A target cell transduced with a viral particle comprising morethan one polypeptide may express each of the polypeptides in a singlecell.

In one embodiment, the payload region may comprise the components asshown in FIG. 1. The payload region 110 is located within the viralgenome 100. At the 5′ and/or the 3′ end of the payload region 110 theremay be at least one inverted terminal repeat (ITR) 120. Within thepayload region, there is a promoter region 130, an intron region 140 anda coding region 150.

Where the AAV particle payload region encodes a polypeptide, thepolypeptide may be a peptide or protein. The viral genomes encodingpolypeptides described herein may be useful in the fields of humandisease, viruses, infections veterinary applications and a variety of invivo and in vitro settings.

In some embodiments, the AAV particles are useful in the field ofmedicine for the treatment, prophylaxis, palliation or amelioration ofneurological diseases and/or disorders.

In some embodiments, the AAV particles are useful in the field ofmedicine for the treatment, prophylaxis, palliation or amelioration ofParkinson's Disease.

In some embodiments, the AAV particles are useful in the field ofmedicine for the treatment, prophylaxis, palliation or amelioration ofdiseases of the central nervous system.

The Nature of the Polypeptides and Variants

Amino acid sequences encoded by payload regions of the viral genomes ofthe disclosure may be translated as a whole polypeptide, a plurality ofpolypeptides or fragments of polypeptides, which independently may beencoded by one or more nucleic acids, fragments of nucleic acids orvariants of any of the aforementioned. As used herein, “polypeptide”means a polymer of amino acid residues (natural or unnatural) linkedtogether most often by peptide bonds. The term, as used herein, refersto proteins, polypeptides, and peptides of any size, structure, orfunction. In some instances, the polypeptide encoded is smaller thanabout 50 amino acids and the polypeptide is then termed a peptide. Ifthe polypeptide is a peptide, it will be at least about 2, 3, 4, or atleast 5 amino acid residues long. Thus, polypeptides include geneproducts, naturally occurring polypeptides, synthetic polypeptides,homologs, orthologs, paralogs, fragments and other equivalents,variants, and analogs of the foregoing. A polypeptide may be a singlemolecule or may be a multi-molecular complex such as a dimer, trimer ortetramer. They may also comprise single chain or multichain polypeptidesand may be associated or linked. The term polypeptide may also apply toamino acid polymers in which one or more amino acid residues are anartificial chemical analogue of a corresponding naturally occurringamino acid.

The term “polypeptide variant” refers to molecules which differ in theiramino acid sequence from a native or reference sequence. The amino acidsequence variants may possess substitutions, deletions, and/orinsertions at certain positions within the amino acid sequence, ascompared to a native or reference sequence. Ordinarily, variants willpossess at least about 50% identity (homology) to a native or referencesequence, and preferably, they will be at least about 80%, morepreferably at least about 90% identical (homologous) to a native orreference sequence.

In some embodiments “variant mimics” are provided. As used herein, theterm “variant mimic” is one which contains one or more amino acids whichwould mimic an activated sequence. For example, glutamate may serve as amimic for phosphoro-threonine and/or phosphoro-serine. Alternatively,variant mimics may result in deactivation or in an inactivated productcontaining the mimic, e.g., phenylalanine may act as an inactivatingsubstitution for tyrosine; or alanine may act as an inactivatingsubstitution for serine.

The term “amino acid sequence variant” refers to molecules with somedifferences in their amino acid sequences as compared to a native orstarting sequence. The amino acid sequence variants may possesssubstitutions, deletions, and/or insertions at certain positions withinthe amino acid sequence. “Native” or “starting” sequence should not beconfused with a wild type sequence. As used herein, a native or startingsequence is a relative term referring to an original molecule againstwhich a comparison may be made. “Native” or “starting” sequences ormolecules may represent the wild-type (that sequence found in nature)but do not have to be the wild-type sequence.

Ordinarily, variants will possess at least about 70% homology to anative sequence, and preferably, they will be at least about 80%, morepreferably at least about 90% homologous to a native sequence.“Homology” as it applies to amino acid sequences is defined as thepercentage of residues in the candidate amino acid sequence that areidentical with the residues in the amino acid sequence of a secondsequence after aligning the sequences and introducing gaps, ifnecessary, to achieve the maximum percent homology. Methods and computerprograms for the alignment are well known in the art. It is understoodthat homology depends on a calculation of percent identity but maydiffer in value due to gaps and penalties introduced in the calculation.

By “homologs” as it applies to amino acid sequences is meant thecorresponding sequence of other species having substantial identity to asecond sequence of a second species.

“Analogs” is meant to include polypeptide variants which differ by oneor more amino acid alterations, e.g., substitutions, additions ordeletions of amino acid residues that still maintain the properties ofthe parent polypeptide.

Sequence tags or amino acids, such as one or more lysines, can be addedto the peptide sequences of the disclosure (e.g., at the N-terminal orC-terminal ends). Sequence tags can be used for peptide purification orlocalization. Lysines can be used to increase peptide solubility or toallow for biotinylation. Alternatively, amino acid residues located atthe carboxy and amino terminal regions of the amino acid sequence of apeptide or protein may optionally be deleted providing for truncatedsequences. Certain amino acids (e.g., C-terminal or N-terminal residues)may alternatively be deleted depending on the use of the sequence, asfor example, expression of the sequence as part of a larger sequencewhich is soluble, or linked to a solid support.

“Substitutional variants” when referring to proteins are those that haveat least one amino acid residue in a native or starting sequence removedand a different amino acid inserted in its place at the same position.The substitutions may be single, where only one amino acid in themolecule has been substituted, or they may be multiple, where two ormore amino acids have been substituted in the same molecule.

As used herein the term “conservative amino acid substitution” refers tothe substitution of an amino acid that is normally present in thesequence with a different amino acid of similar size, charge, orpolarity. Examples of conservative substitutions include thesubstitution of a non-polar (hydrophobic) residue such as isoleucine,valine and leucine for another non-polar residue. Likewise, examples ofconservative substitutions include the substitution of one polar(hydrophilic) residue for another such as between arginine and lysine,between glutamine and asparagine, and between glycine and serine.Additionally, the substitution of a basic residue such as lysine,arginine or histidine for another, or the substitution of one acidicresidue such as aspartic acid or glutamic acid for another acidicresidue are additional examples of conservative substitutions. Examplesof non-conservative substitutions include the substitution of anon-polar (hydrophobic) amino acid residue such as isoleucine, valine,leucine, alanine, methionine for a polar (hydrophilic) residue such ascysteine, glutamine, glutamic acid or lysine and/or a polar residue fora non-polar residue.

“Insertional variants” when referring to proteins are those with one ormore amino acids inserted immediately adjacent to an amino acid at aparticular position in a native or starting sequence. “Immediatelyadjacent” to an amino acid means connected to either the alpha-carboxyor alpha-amino functional group of the amino acid.

“Deletional variants” when referring to proteins, are those with one ormore amino acids in the native or starting amino acid sequence removed.Ordinarily, deletional variants will have one or more amino acidsdeleted in a particular region of the molecule.

As used herein, the term “derivative” is used synonymously with the term“variant” and refers to a molecule that has been modified or changed inany way relative to a reference molecule or starting molecule. In someembodiments, derivatives include native or starting proteins that havebeen modified with an organic proteinaceous or non-proteinaceousderivatizing agent, and post-translational modifications. Covalentmodifications are traditionally introduced by reacting targeted aminoacid residues of the protein with an organic derivatizing agent that iscapable of reacting with selected side-chains or terminal residues, orby harnessing mechanisms of post-translational modifications thatfunction in selected recombinant host cells. The resultant covalentderivatives are useful in programs directed at identifying residuesimportant for biological activity, for immunoassays, or for thepreparation of anti-protein antibodies for immunoaffinity purificationof the recombinant glycoprotein. Such modifications are within theordinary skill in the art and are performed without undueexperimentation.

Certain post-translational modifications are the result of the action ofrecombinant host cells on the expressed polypeptide. Glutaminyl andasparaginyl residues are frequently post-translationally deamidated tothe corresponding glutamyl and aspartyl residues. Alternatively, theseresidues are deamidated under mildly acidic conditions. Either form ofthese residues may be present in the proteins used in accordance withthe present disclosure.

Other post-translational modifications include hydroxylation of prolineand lysine, phosphorylation of hydroxyl groups of seryl or threonylresidues, methylation of the alpha-amino groups of lysine, arginine, andhistidine side chains (T. E. Creighton, Proteins: Structure andMolecular Properties, W.H. Freeman & Co., San Francisco, pp. 79-86(1983)).

“Features” when referring to proteins are defined as distinct amino acidsequence-based components of a molecule. Features of the proteins of thepresent disclosure include surface manifestations, local conformationalshape, folds, loops, half-loops, domains, half-domains, sites, terminior any combination thereof.

As used herein when referring to proteins the term “surfacemanifestation” refers to a polypeptide based component of a proteinappearing on an outermost surface.

As used herein when referring to proteins the term “local conformationalshape” means a polypeptide based structural manifestation of a proteinwhich is located within a definable space of the protein.

As used herein when referring to proteins the term “fold” means theresultant conformation of an amino acid sequence upon energyminimization. A fold may occur at the secondary or tertiary level of thefolding process. Examples of secondary level folds include beta sheetsand alpha helices. Examples of tertiary folds include domains andregions formed due to aggregation or separation of energetic forces.Regions formed in this way include hydrophobic and hydrophilic pockets,and the like.

As used herein the term “turn” as it relates to protein conformationmeans a bend which alters the direction of the backbone of a peptide orpolypeptide and may involve one, two, three or more amino acid residues.

As used herein when referring to proteins the term “loop” refers to astructural feature of a peptide or polypeptide which reverses thedirection of the backbone of a peptide or polypeptide and comprises fouror more amino acid residues. Oliva et al. have identified at least 5classes of protein loops (J. Mol Biol 266 (4): 814-830; 1997).

As used herein when referring to proteins the term “half-loop” refers toa portion of an identified loop having at least half the number of aminoacid residues as the loop from which it is derived. It is understoodthat loops may not always contain an even number of amino acid residues.Therefore, in those cases where a loop contains or is identified tocomprise an odd number of amino acids, a half-loop of the odd-numberedloop will comprise the whole number portion or next whole number portionof the loop (number of amino acids of the loop/2+/−0.5 amino acids). Forexample, a loop identified as a 7 amino acid loop could producehalf-loops of 3 amino acids or 4 amino acids (7/2=3.5+/−0.5 being 3 or4).

As used herein when referring to proteins the term “domain” refers to amotif of a polypeptide having one or more identifiable structural orfunctional characteristics or properties (e.g., binding capacity,serving as a site for protein-protein interactions).

As used herein when referring to proteins the term “half-domain” meansportion of an identified domain having at least half the number of aminoacid residues as the domain from which it is derived. It is understoodthat domains may not always contain an even number of amino acidresidues. Therefore, in those cases where a domain contains or isidentified to comprise an odd number of amino acids, a half-domain ofthe odd-numbered domain will comprise the whole number portion or nextwhole number portion of the domain (number of amino acids of thedomain/2+/−0.5 amino acids). For example, a domain identified as a 7amino acid domain could produce half-domains of 3 amino acids or 4 aminoacids (7/2=3.5+/−0.5 being 3 or 4). It is also understood thatsub-domains may be identified within domains or half-domains, thesesubdomains possessing less than all of the structural or functionalproperties identified in the domains or half domains from which theywere derived. It is also understood that the amino acids that compriseany of the domain types herein need not be contiguous along the backboneof the polypeptide (i.e., nonadjacent amino acids may fold structurallyto produce a domain, half-domain or subdomain).

As used herein when referring to proteins the terms “site” as itpertains to amino acid based embodiments is used synonymous with “aminoacid residue” and “amino acid side chain”. A site represents a positionwithin a peptide or polypeptide that may be modified, manipulated,altered, derivatized or varied within the polypeptide based molecules ofthe present disclosure.

As used herein the terms “termini or terminus” when referring toproteins refers to an extremity of a peptide or polypeptide. Suchextremity is not limited only to the first or final site of the peptideor polypeptide but may include additional amino acids in the terminalregions. The polypeptide based molecules of the present disclosure maybe characterized as having both an N-terminus (terminated by an aminoacid with a free amino group (NH2)) and a C-terminus (terminated by anamino acid with a free carboxyl group (COOH)). Proteins of thedisclosure are in some cases made up of multiple polypeptide chainsbrought together by disulfide bonds or by non-covalent forces(multimers, oligomers). These sorts of proteins will have multiple N-and C-termini. Alternatively, the termini of the polypeptides may bemodified such that they begin or end, as the case may be, with anon-polypeptide based moiety such as an organic conjugate.

Once any of the features have been identified or defined as a componentof a molecule of the disclosure, any of several manipulations and/ormodifications of these features may be performed by moving, swapping,inverting, deleting, randomizing or duplicating. Furthermore, it isunderstood that manipulation of features may result in the same outcomeas a modification to the molecules of the disclosure. For example, amanipulation which involves deleting a domain would result in thealteration of the length of a molecule just as modification of a nucleicacid to encode less than a full length molecule would.

Modifications and manipulations can be accomplished by methods known inthe art such as site directed mutagenesis. The resulting modifiedmolecules may then be tested for activity using in vitro or in vivoassays such as those described herein or any other suitable screeningassay known in the art.

Payload: AADC Polynucleotide Constructs

According to the present disclosure, aromatic L-amino acid decarboxylase(AADC; also known as dopa decarboxylase and DDC) polynucleotides areprovided which function alone or in combination with additional nucleicacid sequence(s) to encode the AADC protein. As used herein an “AADCpolynucleotide” is any nucleic acid polymer which encodes an AADCprotein and when present in a vector, plasmid or translatable construct,expresses such AADC protein in a cell, tissue, organ or organism.

AADC polynucleotides include precursor molecules which are processedinside the cell. AADC polynucleotides or the processed forms thereof maybe encoded in a plasmid, vector, genome or other nucleic acid expressionvector for delivery to a cell.

In some embodiments AADC polynucleotides are designed as components ofAAV viral genomes and packaged in AAV particles which are processedwithin the cell to produce the wild type AADC protein.

In some embodiments, the AADC polynucleotide may be the payload of theAAV particle.

As used herein, the wild type AADC protein may be any of the naturallyoccurring isoforms or variants from the DDC gene. Multiple alternativelyspliced transcript variants encoding different isoforms of AADC havebeen identified. Specifically, the DDC gene produces seven transcriptvariants that encode six distinct isoforms. DDC transcript variants 1and 2 both encode AADC isoform 1. In some embodiments, the AADCpolynucleotides encode DDC transcript variant 2, thereby encoding anative AADC isoform 1 (NCBI Reference Sequence: NP 000781.1). Thissequence is given here:

(SEQ ID NO: 978) MNASEFRRRGKEMVDYVANYMEGIEGRQVYPDVEPGYLRPLIPAAAPQEPDTFEDIINDVEKIIMPGVTHWHSPYFFAYFPTASSYPAMLADMLCGAIGCIGFSWAASPACTELETVMMDWLGKMLELPKAFLNEKAGEGGGVIQGSASEATLVALLAARTKVIHRLQAASPELTQAAIMEKLVAYSSDQAHSSVERAGLIGGVKLKAIPSDGNFAMRASALQEALERDKAAGLIPFFMVATLGTTTCCSFDNLLEVGPICNKEDIWLHVDAAYAGSAFICPEFRHLLNGVEFADSFNFNPHKWLLVNFDCSAMWVKKRTDLTGAFRLDPTYLKHSHQDSGLITDYRHWQIPLGRRFRSLKMWFVFRMYGVKGLQAYIRKHVQLSHEFESLVRQDPRFEICVEVILGLVCFRLKGSNKVNEALLQRINSAKKIHLVPCHLRDKFVLRFAICSRTVESAHVQRAWEHIKELAADVLRAERE

The AADC polynucleotides of the disclosure, may be engineered to containmodular elements and/or sequence motifs assembled to create AADCpolynucleotide constructs.

According to the present disclosure, AADC polynucleotides are provided.Such polynucleotides comprise nucleic acid polymers which comprise aregion of linked nucleosides encoding one or more isoforms or variantsof the AADC protein.

In some embodiments, the AADC polynucleotide comprises a codon optimizedtranscript encoding an AADC protein.

In some embodiments, the AADC polynucleotide comprises a sequence regionencoding one or more wild type isoforms or variants of an AADC protein.Such polynucleotides may also comprise a sequence region encoding anyone or more of the following: a 5′ ITR, a cytomegalovirus (CMV)Enhancer, a CMV Promoter, an ie1 exon 1, an ie1 intron1, an hbBglobinintron2, an hBglobin exon 3, a 5′ UTR, a 3′ UTR, an hGH poly(A) signal,and/or a 3′ ITR. Such sequence regions are taught herein or may be anyof those known in the art.

In some embodiments, the AADC polynucleotide comprises a SEQ ID NO: 979or a fragment or variant thereof.

In one embodiment, the AADC polynucleotide comprises a sequence whichhas a percent identity to any of SEQ ID NO: 979 or a fragment or variantthereof. The AADC polynucleotide may have 1%, 2%, 3%, 4%, 5%, 6%, 7%,8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,75%, 80%, 85%, 90%, 95%, 99% or 100% identity to any of SEQ ID NO: 979or a fragment or variant thereof. The AADC polynucleotide may have1-10%, 10-20%, 30-40%, 50-60%, 50-70%, 50-80%, 50-90%, 50-99%, 50-100%,60-70%, 60-80%, 60-90%, 60-99%, 60-100%, 70-80%, 70-90%, 70-99%,70-100%, 80-85%, 80-90%, 80-95%, 80-99%, 80-100%, 90-95%, 90-99%, or90-100% to any of SEQ ID NO: 979 or a fragment or variant thereof. As anon-limiting example, the AADC polynucleotide comprises a sequence whichas 80% identity to any of SEQ ID NO: 979 or a fragment or variantthereof. As another non-limiting example, the AADC polynucleotidecomprises a sequence which as 85% identity to any of SEQ ID NO: 979 or afragment or variant thereof. As another non-limiting example, the AADCpolynucleotide comprises a sequence which as 90% identity to any of SEQID NO: 979 or a fragment or variant thereof. As another non-limitingexample, the AADC polynucleotide comprises a sequence which as 95%identity to any of SEQ ID NO: 979 or a fragment or variant thereof. Asanother non-limiting example, the AADC polynucleotide comprises asequence which as 99% identity to any of SEQ ID NO: 979 or a fragment orvariant thereof.

In some embodiments, the coding region of the AADC polynucleotide is1440 nucleotides in length. Such an AADC polynucleotide may be codonoptimized over all or a portion of the polynucleotide.

In some embodiments, the AADC polynucleotide comprises any of SEQ ID NO:979 or a fragment or variant thereof but lacking the 5′ and/or 3′ ITRs.Such a polynucleotide may be incorporated into a plasmid or vector andutilized to express the encoded AADC protein.

In one embodiment, the AADC polynucleotides may be produced in insectcells (e.g., SD cells).

In one embodiment, the AADC polynucleotides may be produced using tripletransfection.

In one embodiment, the AADC polynucleotide may comprise a codonoptimized open reading frame of an AADC mRNA, at least one 5′ITR and atleast one 3′UTR where the one or more of the 5′ITRs may be located atthe 5′ end of the promoter region and one or more 3′ ITRs may be locatedat the 3′ end of the poly(A) signal. The AADC mRNA may comprise apromoter region, a 5′untranslated region (UTR), a 3′UTR and a poly(A)signal. The promoter region may include, but is not limited to, enhancerelement, a promoter element, a first exon region, a first intron region,a second intron region and a second exon region. As a non-limitingexample, the enhancer element and the promoter element are derived fromCMV. As another non-limiting example, the first exon region is ie1 exon1 or fragments thereof, the first intron region is ie1 intron 1 orfragments thereof, the second intron region is hbBglobin intron 2 orfragments thereof and the second exon region is hbBglobin exon 3 orfragments thereof. As yet another non-limiting example, the poly(A)signal is derived from human growth hormone.

In one embodiment, at least one element may be used with the AADCpolynucleotides described herein to enhance the transgene targetspecificity and expression (See e.g., Powell et al. Viral ExpressionCassette Elements to Enhance Transgene Target Specificity and Expressionin Gene Therapy, 2015; the contents of which are herein incorporated byreference in its entirety).Non-limiting examples of elements to enhancethe transgene target specificity and expression include promoters,endogenous miRNAs, post-transcriptional regulatory elements (PREs),polyadenylation (PolyA) signal sequences and upstream enhancers (USEs),CMV enhancers and introns.

In one embodiment, at least one element may be used with the AADCpolynucleotides described herein to enhance the transgene targetspecificity and expression (See e.g., Powell et al. Viral ExpressionCassette Elements to Enhance Transgene Target Specificity and Expressionin Gene Therapy, 2015; the contents of which are herein incorporated byreference in its entirety) such as promoters.

In one embodiment, the AADC polynucleotide is encoded in a plasmid orvector, which may be derived from an adeno-associated virus (AAV). TheAAV may comprise a capsid serotype such as, but not limited to, PHP.B,PHP.A, AAV1, AAV2, AAV2G9, AAV3, AAV3a, AAV3b, AAV3-3, AAV4, AAV4-4,AAV5, AAV6, AAV6.1, AAV6.2, AAV6.1.2, AAV7, AAV7.2, AAV8, AAV9, AAV9.11,AAV9.13, AAV9.16, AAV9.24, AAV9.45, AAV9.47, AAV9.61, AAV9.68, AAV9.84,AAV9.9, AAV10, AAV11, AAV12, AAV16.3, AAV24.1, AAV27.3, AAV42.12,AAV42-1b, AAV42-2, AAV42-3a, AAV42-3b, AAV42-4, AAV42-5a, AAV42-5b,AAV42-6b, AAV42-8, AAV42-10, AAV42-11, AAV42-12, AAV42-13, AAV42-15,AAV42-aa, AAV43-1, AAV43-12, AAV43-20, AAV43-21, AAV43-23, AAV43-25,AAV43-5, AAV44.1, AAV44.2, AAV44.5, AAV223.1, AAV223.2, AAV223.4,AAV223.5, AAV223.6, AAV223.7, AAV1-7/rh.48, AAV1-8/rh.49, AAV2-15/rh.62,AAV2-3/rh.61, AAV2-4/rh.50, AAV2-5/rh.51, AAV3.1/hu.6, AAV3.1/hu.9,AAV3-9/rh.52, AAV3-11/rh.53, AAV4-8/r11.64, AAV4-9/rh.54, AAV4-19/rh.55,AAV5-3/rh.57, AAV5-22/rh.58, AAV7.3/hu.7, AAV16.8/hu.10, AAV16.12/hu.11,AAV29.3/bb.1, AAV29.5/bb.2, AAV106.1/hu.37, AAV114.3/hu.40,AAV127.2/hu.41, AAV127.5/hu.42, AAV128.3/hu.44, AAV130.4/hu.48,AAV145.1/hu.53, AAV145.5/hu.54, AAV145.6/hu.55, AAV161.10/hu.60,AAV161.6/hu.61, AAV33.12/hu.17, AAV33.4/hu.15, AAV33.8/hu.16,AAV52/hu.19, AAV52.1/hu.20, AAV58.2/hu.25, AAVA3.3, AAVA3.4, AAVA3.5,AAVA3.7, AAVC1, AAVC2, AAVC5, AAV-DJ, AAV-DJ8, AAVF3, AAVF5, AAVH2,AAVrh.72, AAVhu.8, AAVrh.68, AAVrh.70, AAVpi.1, AAVpi.3, AAVpi.2,AAVrh.60, AAVrh.44, AAVrh.65, AAVrh.55, AAVrh.47, AAVrh.69, AAVrh.45,AAVrh.59, AAVhu.12, AAVH6, AAVLK03, AAVH-1/hu.1, AAVH-5/hu.3,AAVLG-10/rh.40, AAVLG-4/rh.38, AAVLG-9/hu.39, AAVN721-8/rh.43, AAVCh.5,AAVCh.5R1, AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5, AAVCy.5R1, AAVCy.5R2,AAVCy.5R3, AAVCy.5R4, AAVcy.6, AAVhu.1, AAVhu.2, AAVhu.3, AAVhu.4,AAVhu.5, AAVhu.6, AAVhu.7, AAVhu.9, AAVhu.10, AAVhu.11, AAVhu.13,AAVhu.15, AAVhu.16, AAVhu.17, AAVhu.18, AAVhu.20, AAVhu.21, AAVhu.22,AAVhu.23.2, AAVhu.24, AAVhu.25, AAVhu.27, AAVhu.28, AAVhu.29, AAVhu.29R,AAVhu.31, AAVhu.32, AAVhu.34, AAVhu.35, AAVhu.37, AAVhu.39, AAVhu.40,AAVhu.41, AAVhu.42, AAVhu.43, AAVhu.44, AAVhu.44R1, AAVhu.44R2,AAVhu.44R3, AAVhu.45, AAVhu.46, AAVhu.47, AAVhu.48, AAVhu.48R1,AAVhu.48R2, AAVhu.48R3, AAVhu.49, AAVhu.51, AAVhu.52, AAVhu.54,AAVhu.55, AAVhu.56, AAVhu.57, AAVhu.58, AAVhu.60, AAVhu.61, AAVhu.63,AAVhu.64, AAVhu.66, AAVhu.67, AAVhu.14/9, AAVhu.t 19, AAVrh.2, AAVrh.2R,AAVrh.8, AAVrh.8R, AAVrh.10, AAVrh.12, AAVrh.13, AAVrh.13R, AAVrh.14,AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.20, AAVrh.21, AAVrh.22, AAVrh.23,AAVrh.24, AAVrh.25, AAVrh.31, AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35,AAVrh.36, AAVrh.37, AAVrh.37R2, AAVrh.38, AAVrh.39, AAVrh.40, AAVrh.46,AAVrh.48, AAVrh.48.1, AAVrh.48.1.2, AAVrh.48.2, AAVrh.49, AAVrh.51,AAVrh.52, AAVrh.53, AAVrh.54, AAVrh.56, AAVrh.57, AAVrh.58, AAVrh.61,AAVrh.64, AAVrh.64R1, AAVrh.64R2, AAVrh.67, AAVrh.73, AAVrh.74,AAVrh8R,AAVrh8R A586R mutant, AAVrh8R R533A mutant, AAAV, BAAV, caprine AAV,bovine AAV, AAVhE1.1, AAVhEr1.5, AAVhER1.14, AAVhEr1.8, AAVhEr1.16,AAVhEr1.18, AAVhEr1.35, AAVhEr1.7, AAVhEr1.36, AAVhEr2.29, AAVhEr2.4,AAVhEr2.16, AAVhEr2.30, AAVhEr2.31, AAVhEr2.36, AAVhER1.23, AAVhEr3.1,AAV2.5T, AAV-PAEC, AAV-LK01, AAV-LK02, AAV-LK03, AAV-LK04, AAV-LK05,AAV-LK06, AAV-LK07, AAV-LK08, AAV-LK09, AAV-LK10, AAV-LK11, AAV-LK12,AAV-LK13, AAV-LK14, AAV-LK15, AAV-LK16, AAV-LK17, AAV-LK18, AAV-LK19,AAV-PAEC2, AAV-PAEC4, AAV-PAEC6, AAV-PAEC7, AAV-PAEC8, AAV-PAEC11,AAV-PAEC12, AAV-2-pre-miRNA-101, AAV-8h, AAV-8b, AAV-h, AAV-b, AAV SM10-2, AAV Shuffle 100-1, AAV Shuffle 100-3, AAV Shuffle 100-7, AAVShuffle 10-2, AAV Shuffle 10-6, AAV Shuffle 10-8, AAV Shuffle 100-2, AAVSM 10-1, AAV SM 10-8, AAV SM 100-3, AAV SM 100-10, BNP61 AAV, BNP62 AAV,BNP63 AAV, AAVrh.50, AAVrh.43, AAVrh.62, AAVrh.48, AAVhu.19, AAVhu.11,AAVhu.53, AAV4-8/rh.64, AAVLG-9/hu.39, AAV54.5/hu.23, AAV54.2/hu.22,AAV54.7/hu.24, AAV54.1/hu.21, AAV54.4R/hu.27, AAV46.2/hu.28,AAV46.6/hu.29, AAV128.1/hu.43, true type AAV (ttAAV), UPENN AAV 10,Japanese AAV 10 serotypes, AAV CBr-7.1, AAV CBr-7.10, AAV CBr-7.2, AAVCBr-7.3, AAV CBr-7.4, AAV CBr-7.5, AAV CBr-7.7, AAV CBr-7.8, AAVCBr-B7.3, AAV CBr-B7.4, AAV CBr-E1, AAV CBr-E2, AAV CBr-E3, AAV CBr-E4,AAV CBr-E5, AAV CBr-e5, AAV CBr-E6, AAV CBr-E7, AAV CBr-E8, AAV CHt-1,AAV CHt-2, AAV CHt-3, AAV CHt-6.1, AAV CHt-6.10, AAV CHt-6.5, AAVCHt-6.6, AAV CHt-6.7, AAV CHt-6.8, AAV CHt-P1, AAV CHt-P2, AAV CHt-P5,AAV CHt-P6, AAV CHt-P8, AAV CHt-P9, AAV CKd-1, AAV CKd-10, AAV CKd-2,AAV CKd-3, AAV CKd-4, AAV CKd-6, AAV CKd-7, AAV CKd-8, AAV CKd-B1, AAVCKd-B2, AAV CKd-B3, AAV CKd-B4, AAV CKd-B5, AAV CKd-B6, AAV CKd-B7, AAVCKd-B8, AAV CKd-H1, AAV CKd-H2, AAV CKd-H3, AAV CKd-H4, AAV CKd-H5, AAVCKd-H6, AAV CKd-N3, AAV CKd-N4, AAV CKd-N9, AAV CLg-F1, AAV CLg-F2, AAVCLg-F3, AAV CLg-F4, AAV CLg-F5, AAV CLg-F6, AAV CLg-F7, AAV CLg-F8, AAVCLv-1, AAV CLv1-1, AAV Clv1-10, AAV CLv1-2, AAV CLv-12, AAV CLv1-3, AAVCLv-13, AAV CLv1-4, AAV Clv1-7, AAV Clv1-8, AAV Clv1-9, AAV CLv-2, AAVCLv-3, AAV CLv-4, AAV CLv-6, AAV CLv-8, AAV CLv-D1, AAV CLv-D2, AAVCLv-D3, AAV CLv-D4, AAV CLv-D5, AAV CLv-D6, AAV CLv-D7, AAV CLv-D8, AAVCLv-E1, AAV CLv-K1, AAV CLv-K3, AAV CLv-K6, AAV CLv-L4, AAV CLv-L5, AAVCLv-L6, AAV CLv-M1, AAV CLv-M11, AAV CLv-M2, AAV CLv-M5, AAV CLv-M6, AAVCLv-M7, AAV CLv-M8, AAV CLv-M9, AAV CLv-R1, AAV CLv-R2, AAV CLv-R3, AAVCLv-R4, AAV CLv-R5, AAV CLv-R6, AAV CLv-R7, AAV CLv-R8, AAV CLv-R9, AAVCSp-1, AAV CSp-10, AAV CSp-11, AAV CSp-2, AAV CSp-3, AAV CSp-4, AAVCSp-6, AAV CSp-7, AAV CSp-8, AAV CSp-8.10, AAV CSp-8.2, AAV CSp-8.4, AAVCSp-8.5, AAV CSp-8.6, AAV CSp-8.7, AAV CSp-8.8, AAV CSp-8.9, AAV CSp-9,AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5, AAVF1/HSC1, AAVF11/HSC11,AAVF12/HSC12, AAVF13/HSC13, AAVF14/HSC14, AAVF15/HSC15, AAVF16/HSC16,AAVF17/HSC17, AAVF2/HSC2, AAVF3/HSC3, AAVF4/HSC4, AAVF5/HSC5,AAVF6/HSC6, AAVF7/HSC7, AAVF8/HSC8, AAVF9/HSC9, PHP.B (AAV-PHP.B), PHP.A(AAV.PHP.A), G2B-26, G2B-13, TH1.1-32, TH1.1-35, AAVPHP.B2, AAVPHP.B3,AAVPHP.N/PHP.B-DGT, AAVPHP.B-EST, AAVPHP.B-GGT, AAVPHP.B-ATP,AAVPHP.B-ATT-T, AAVPHP.B-DGT-T, AAVPHP.B-GGT-T, AAVPHP.B-SGS,AAVPHP.B-AQP, AAVPHP.B-QQP, AAVPHP.B-SNP(3), AAVPHP.B-SNP, AAVPHP.B-QGT,AAVPHP.B-NQT, AAVPHP.B-EGS, AAVPHP.B-SGN, AAVPHP.B-EGT, AAVPHP.B-DST,AAVPHP.B-DST, AAVPHP.B-STP, AAVPHP.B-PQP, AAVPHP.B-SQP, AAVPHP.B-QLP,AAVPHP.B-TMP, AAVPHP.B-TTP, AAVPHP.S/G2A12, AAVG2A15/G2A3, AAVG2B4,and/or AAVG2B5, and variants thereof.

II. Formulation and Delivery Pharmaceutical Compositions

According to the present disclosure the AAV particles may be prepared aspharmaceutical compositions. It will be understood that suchcompositions necessarily comprise one or more active ingredients and,most often, a pharmaceutically acceptable excipient.

Relative amounts of the active ingredient (e.g. AAV particle), apharmaceutically acceptable excipient, and/or any additional ingredientsin a pharmaceutical composition in accordance with the presentdisclosure may vary, depending upon the identity, size, and/or conditionof the subject being treated and further depending upon the route bywhich the composition is to be administered. For example, thecomposition may comprise between 0.1% and 99% (w/w) of the activeingredient. By way of example, the composition may comprise between 0.1%and 100%, e.g., between 0.5 and 50%, between 1-30%, between 5-80%, atleast 80% (w/w) active ingredient. In some embodiments, the AAV particlepharmaceutical compositions described herein may comprise at least onepayload. As a non-limiting example, the pharmaceutical compositions maycontain an AAV particle with 1, 2, 3, 4 or 5 payloads.

Although the descriptions of pharmaceutical compositions provided hereinare principally directed to pharmaceutical compositions which aresuitable for administration to humans, it will be understood by theskilled artisan that such compositions are generally suitable foradministration to any other animal, e.g., to non-human animals, e.g.non-human mammals.

Modification of pharmaceutical compositions suitable for administrationto humans in order to render the compositions suitable foradministration to various animals is well understood, and the ordinarilyskilled veterinary pharmacologist can design and/or perform suchmodification with merely ordinary, if any, experimentation. Subjects towhich administration of the pharmaceutical compositions is contemplatedinclude, but are not limited to, humans and/or other primates; mammals,including commercially relevant mammals such as cattle, pigs, horses,sheep, cats, dogs, mice, rats, birds, including commercially relevantbirds such as poultry, chickens, ducks, geese, and/or turkeys.

In some embodiments, compositions are administered to humans, humanpatients or subjects.

Formulations

Formulations of the present disclosure can include, without limitation,saline, liposomes, lipid nanoparticles, polymers, peptides, proteins,cells transfected with AAV particles (e.g., for transfer ortransplantation into a subject) and combinations thereof.

Formulations of the pharmaceutical compositions described herein may beprepared by any method known or hereafter developed in the art ofpharmacology. As used herein the term “pharmaceutical composition”refers to compositions comprising at least one active ingredient andoptionally one or more pharmaceutically acceptable excipients.

In general, such preparatory methods include the step of associating theactive ingredient with an excipient and/or one or more other accessoryingredients. As used herein, the phrase “active ingredient” generallyrefers either to an AAV particle carrying a payload region encoding thepolypeptides of the disclosure or to the end product encoded by a viralgenome of by an AAV particle as described herein.

Formulations of the AAV particles and pharmaceutical compositionsdescribed herein may be prepared by any method known or hereafterdeveloped in the art of pharmacology. In general, such preparatorymethods include the step of bringing the active ingredient intoassociation with an excipient and/or one or more other accessoryingredients, and then, if necessary and/or desirable, dividing, shapingand/or packaging the product into a desired single- or multi-dose unit.

A pharmaceutical composition in accordance with the present disclosuremay be prepared, packaged, and/or sold in bulk, as a single unit dose,and/or as a plurality of single unit doses. As used herein, a “unitdose” refers to a discrete amount of the pharmaceutical compositioncomprising a predetermined amount of the active ingredient. The amountof the active ingredient is generally equal to the dosage of the activeingredient which would be administered to a subject and/or a convenientfraction of such a dosage such as, for example, one-half or one-third ofsuch a dosage.

In one embodiment, the AAV particles of the disclosure may be formulatedin PBS with 0.001% of pluronic acid (F-68) at a pH of about 7.0.

In some embodiments, the AAV formulations described herein may containsufficient AAV particles for expression of at least one expressedfunctional payload. As a non-limiting example, the AAV particles maycontain viral genomes encoding 1, 2, 3, 4 or 5 functional payloads.

According to the present disclosure AAV particles may be formulated forCNS delivery. Agents that cross the brain blood barrier may be used. Forexample, some cell penetrating peptides that can target molecules to thebrain blood barrier endothelium may be used for formulation (e.g.,Mathupala, Expert Opin Ther Pat., 2009, 19, 137-140; the content ofwhich is incorporated herein by reference in its entirety).

Excipients and Diluents

The AAV particles of the disclosure can be formulated using one or moreexcipients or diluents to (1) increase stability; (2) increase celltransfection or transduction; (3) permit the sustained or delayedrelease of the payload; (4) alter the biodistribution (e.g., target theviral particle to specific tissues or cell types); (5) increase thetranslation of encoded protein; (6) alter the release profile of encodedprotein and/or (7) allow for regulatable expression of the payload ofthe disclosure.

In some embodiments, a pharmaceutically acceptable excipient may be atleast 95%, at least 96%, at least 97%, at least 98%, at least 99%, or100% pure. In some embodiments, an excipient is approved for use forhumans and for veterinary use. In some embodiments, an excipient may beapproved by United States Food and Drug Administration. In someembodiments, an excipient may be of pharmaceutical grade. In someembodiments, an excipient may meet the standards of the United StatesPharmacopoeia (USP), the European Pharmacopoeia (EP), the BritishPharmacopoeia, and/or the International Pharmacopoeia.

Excipients, as used herein, include, but are not limited to, any and allsolvents, dispersion media, diluents, or other liquid vehicles,dispersion or suspension aids, surface active agents, isotonic agents,thickening or emulsifying agents, preservatives, and the like, as suitedto the particular dosage form desired. Various excipients forformulating pharmaceutical compositions and techniques for preparing thecomposition are known in the art (see Remington: The Science andPractice of Pharmacy, 21st Edition, A. R. Gennaro, Lippincott, Williams& Wilkins, Baltimore, Md., 2006; incorporated herein by reference in itsentirety). The use of a conventional excipient medium may becontemplated within the scope of the present disclosure, except insofaras any conventional excipient medium may be incompatible with asubstance or its derivatives, such as by producing any undesirablebiological effect or otherwise interacting in a deleterious manner withany other component(s) of the pharmaceutical composition.

Exemplary diluents include, but are not limited to, calcium carbonate,sodium carbonate, calcium phosphate, dicalcium phosphate, calciumsulfate, calcium hydrogen phosphate, sodium phosphate lactose, sucrose,cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol,inositol, sodium chloride, dry starch, cornstarch, powdered sugar, etc.,and/or combinations thereof.

In one embodiment, the AAV particles may be formulated in a hydrogelprior to administration. Hydrogels have a degree of flexibility which issimilar to natural tissue as a result of their significant watercontent.

In another embodiment, a hydrogel may be administered to a subject priorto the administration of an AAV particle formulation. As a non-limitingexample, the site of administration of the hydrogel may be within 3inches (e.g., within 2.9, 2.8, 2.7, 2.6, 2.5, 2.4, 2.3, 2.2., 2.1, 2.0,1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6,0.5, 0.4, 0.3, 0.2, 0.1 or less than 0.1 inches) of the site ofadministration of the AAV particle formulation.

Inactive Ingredients

In some embodiments, AAV particle formulations may comprise at least oneinactive ingredient. As used herein, the term “inactive ingredient”refers to one or more agents that do not contribute to the activity ofthe active ingredient of the pharmaceutical composition included informulations. In some embodiments, all, none or some of the inactiveingredients which may be used in the formulations of the presentdisclosure may be approved by the US Food and Drug Administration (FDA).

In one embodiment, the AAV particle pharmaceutical compositions compriseat least one inactive ingredient such as, but not limited to,1,2,6-Hexanetriol;1,2-Dimyristoyl-Sn-Glycero-3-(Phospho-S-(1-Glycerol));1,2-Dimyristoyl-Sn-Glycero-3-Phosphocholine;1,2-Dioleoyl-Sn-Glycero-3-Phosphocholine;1,2-Dipalmitoyl-Sn-Glycero-3-(Phospho-Rac-(1-Glycerol));1,2-Distearoyl-Sn-Glycero-3-(Phospho-Rac-(1-Glycerol));1,2-Distearoyl-Sn-Glycero-3-Phosphocholine; 1-O-Tolylbiguanide;2-Ethyl-1,6-Hexanediol; Acetic Acid; Acetic Acid, Glacial; AceticAnhydride; Acetone; Acetone Sodium Bisulfite; Acetylated LanolinAlcohols; Acetylated Monoglycerides; Acetylcysteine; Acetyltryptophan,DL-; Acrylates Copolymer; Acrylic Acid-Isooctyl Acrylate Copolymer;Acrylic Adhesive 788; Activated Charcoal; Adcote 72A103; Adhesive Tape;Adipic Acid; Aerotex Resin 3730; Alanine; Albumin Aggregated; AlbuminColloidal; Albumin Human; Alcohol; Alcohol, Dehydrated; Alcohol,Denatured; Alcohol, Diluted; Alfadex; Alginic Acid; Alkyl AmmoniumSulfonic Acid Betaine; Alkyl Aryl Sodium Sulfonate; Allantoin; Allyl.Alpha.-Ionone; Almond Oil; Alpha-Terpineol; Alpha-Tocopherol;Alpha-Tocopherol Acetate, Dl-; Alpha-Tocopherol, Dl-; Aluminum Acetate;Aluminum Chlorhydroxy Allantoinate; Aluminum Hydroxide; AluminumHydroxide-Sucrose, Hydrated; Aluminum Hydroxide Gel; Aluminum HydroxideGel F 500; Aluminum Hydroxide Gel F 5000; Aluminum Monostearate;Aluminum Oxide; Aluminum Polyester; Aluminum Silicate; Aluminum StarchOctenylsuccinate; Aluminum Stearate; Aluminum Subacetate; AluminumSulfate Anhydrous; Amerchol C; Amerchol-Cab; Aminomethylpropanol;Ammonia; Ammonia Solution; Ammonia Solution, Strong; Ammonium Acetate;Ammonium Hydroxide; Ammonium Lauryl Sulfate; Ammonium Nonoxynol-4Sulfate; Ammonium Salt Of C-12-C-15 Linear Primary Alcohol Ethoxylate;Ammonium Sulfate; Ammonyx; Amphoteric-2; Amphoteric-9; Anethole;Anhydrous Citric Acid; Anhydrous Dextrose; Anhydrous Lactose; AnhydrousTrisodium Citrate; Aniseed Oil; Anoxid Sbn; Antifoam; Antipyrine;Apaflurane; Apricot Kernel Oil Peg-6 Esters; Aquaphor; Arginine;Arlacel; Ascorbic Acid; Ascorbyl Palmitate; Aspartic Acid; Balsam Peru;Barium Sulfate; Beeswax; Beeswax, Synthetic; Beheneth-10; Bentonite;Benzalkonium Chloride; Benzenesulfonic Acid; Benzethonium Chloride;Benzododecinium Bromide; Benzoic Acid; Benzyl Alcohol; Benzyl Benzoate;Benzyl Chloride; Betadex; Bibapcitide; Bismuth Subgallate; Boric Acid;Brocrinat; Butane; Butyl Alcohol; Butyl Ester Of Vinyl MethylEther/Maleic Anhydride Copolymer (125000 Mw); Butyl Stearate; ButylatedHydroxyanisole; Butylated Hydroxytoluene; Butylene Glycol; Butylparaben;Butyric Acid; C20-40 Pareth-24; Caffeine; Calcium; Calcium Carbonate;Calcium Chloride; Calcium Gluceptate; Calcium Hydroxide; CalciumLactate; Calcobutrol; Caldiamide Sodium; Caloxetate Trisodium;Calteridol Calcium; Canada Balsam; Caprylic/Capric Triglyceride;Caprylic/Capric/Stearic Triglyceride; Captan; Captisol; Caramel;Carbomer 1342; Carbomer 1382; Carbomer 934; Carbomer 934p; Carbomer 940;Carbomer 941; Carbomer 980; Carbomer 981; Carbomer Homopolymer Type B(Allyl Pentaerythritol Crosslinked); Carbomer Homopolymer Type C (AllylPentaerythritol Crosslinked); Carbon Dioxide; Carboxy Vinyl Copolymer;Carboxymethylcellulose; Carboxymethylcellulose Sodium;Carboxypolymethylene; Carrageenan; Carrageenan Salt; Castor Oil; CedarLeaf Oil; Cellulose; Cellulose, Microcrystalline; Cerasynt-Se; Ceresin;Ceteareth-12; Ceteareth-15; Ceteareth-30; Cetearyl Alcohol/Ceteareth-20;Cetearyl Ethylhexanoate; Ceteth-10; Ceteth-2; Ceteth-20; Ceteth-23;Cetostearyl Alcohol; Cetrimonium Chloride; Cetyl Alcohol; Cetyl EstersWax; Cetyl Palmitate; Cetylpyridinium Chloride; Chlorobutanol;Chlorobutanol Hemihydrate; Chlorobutanol, Anhydrous; Chlorocresol;Chloroxylenol; Cholesterol; Choleth; Choleth-24; Citrate; Citric Acid;Citric Acid Monohydrate; Citric Acid, Hydrous; Cocamide Ether Sulfate;Cocamine Oxide; Coco Betaine; Coco Diethanolamide; CocoMonoethanolamide; Cocoa Butter; Coco-Glycerides; Coconut Oil; CoconutOil, Hydrogenated; Coconut Oil/Palm Kernel Oil Glycerides, Hydrogenated;Cocoyl Caprylocaprate; Cola Nitida Seed Extract; Collagen; ColoringSuspension; Corn Oil; Cottonseed Oil; Cream Base; Creatine; Creatinine;Cresol; Croscarmellose Sodium; Crospovidone; Cupric Sulfate; CupricSulfate Anhydrous; Cyclomethicone; Cyclomethicone/Dimethicone Copolyol;Cysteine; Cysteine Hydrochloride; Cysteine Hydrochloride Anhydrous;Cysteine, Dl-; D&C Red No. 28; D&C Red No. 33; D&C Red No. 36; D&C RedNo. 39; D&C Yellow No. 10; Dalfampridine; Daubert 1-5 Pestr (Matte)164z; Decyl Methyl Sulfoxide; Dehydag Wax Sx; Dehydroacetic Acid;Dehymuls E; Denatonium Benzoate; Deoxycholic Acid; Dextran; Dextran 40;Dextrin; Dextrose; Dextrose Monohydrate; Dextrose Solution; DiatrizoicAcid; Diazolidinyl Urea; Dichlorobenzyl Alcohol;Dichlorodifluoromethane; Dichlorotetrafluoroethane; Diethanolamine;Diethyl Pyrocarbonate; Diethyl Sebacate; Diethylene Glycol MonoethylEther; Diethylhexyl Phthalate; Dihydroxyaluminum Aminoacetate;Diisopropanolamine; Diisopropyl Adipate; Diisopropyl Dilinoleate;Dimethicone 350; Dimethicone Copolyol; Dimethicone Mdx4-4210;Dimethicone Medical Fluid 360; Dimethyl Isosorbide; Dimethyl Sulfoxide;Dimethylaminoethyl Methacrylate-Butyl Methacrylate-Methyl MethacrylateCopolymer; Dimethyldioctadecylammonium Bentonite;Dimethylsiloxane/Methylvinylsiloxane Copolymer; Dinoseb Ammonium Salt;Dipalmitoylphosphatidylglycerol, Dl-; Dipropylene Glycol; DisodiumCocoamphodiacetate; Disodium Laureth Sulfosuccinate; Disodium LaurylSulfosuccinate; Disodium Sulfosalicylate; Disofenin; DivinylbenzeneStyrene Copolymer; Dmdm Hydantoin; Docosanol; Docusate Sodium; Duro-Tak280-2516; Duro-Tak 387-2516; Duro-Tak 80-1196; Duro-Tak 87-2070;Duro-Tak 87-2194; Duro-Tak 87-2287; Duro-Tak 87-2296; Duro-Tak 87-2888;Duro-Tak 87-2979; Edetate Calcium Disodium; Edetate Disodium; EdetateDisodium Anhydrous; Edetate Sodium; Edetic Acid; Egg Phospholipids;Entsufon; Entsufon Sodium; Epilactose; Epitetracycline Hydrochloride;Essence Bouquet 9200; Ethanolamine Hydrochloride; Ethyl Acetate; EthylOleate; Ethylcelluloses; Ethylene Glycol; Ethylene Vinyl AcetateCopolymer; Ethylenediamine; Ethylenediamine Dihydrochloride;Ethylene-Propylene Copolymer; Ethylene-Vinyl Acetate Copolymer (28%Vinyl Acetate); Ethylene-Vinyl Acetate Copolymer (9% Vinylacetate);Ethylhexyl Hydroxystearate; Ethylparaben; Eucalyptol; Exametazime; Fat,Edible; Fat, Hard; Fatty Acid Esters; Fatty Acid Pentaerythriol Ester;Fatty Acids; Fatty Alcohol Citrate; Fatty Alcohols; Fd&C Blue No. 1;Fd&C Green No. 3; Fd&C Red No. 4; Fd&C Red No. 40; Fd&C Yellow No. 10(Delisted); Fd&C Yellow No. 5; Fd&C Yellow No. 6; Ferric Chloride;Ferric Oxide; Flavor 89-186; Flavor 89-259; Flavor Df-119; FlavorDf-1530; Flavor Enhancer; Flavor Fig 827118; Flavor Raspberry Pfc-8407;Flavor Rhodia Pharmaceutical No. Rf 451; Fluorochlorohydrocarbons;Formaldehyde; Formaldehyde Solution; Fractionated Coconut Oil; Fragrance3949-5; Fragrance 520a; Fragrance 6.007; Fragrance 91-122; Fragrance9128-Y; Fragrance 93498g; Fragrance Balsam Pine No. 5124; FragranceBouquet 10328; Fragrance Chemoderm 6401-B; Fragrance Chemoderm 6411;Fragrance Cream No. 73457; Fragrance Cs-28197; Fragrance Felton 066m;Fragrance Firmenich 47373; Fragrance Givaudan Ess 9090/1c; FragranceH-6540; Fragrance Herbal 10396; Fragrance Nj-1085; Fragrance P O F1-147;Fragrance Pa 52805; Fragrance Pera Derm D; Fragrance Rbd-9819; FragranceShaw Mudge U-7776; Fragrance Tf 044078; Fragrance Ungerer Honeysuckle K2771; Fragrance Ungerer N5195; Fructose; Gadolinium Oxide; Galactose;Gamma Cyclodextrin; Gelatin; Gelatin, Crosslinked; Gelfoam Sponge;Gellan Gum (Low Acyl); Gelva 737; Gentisic Acid; Gentisic AcidEthanolamide; Gluceptate Sodium; Gluceptate Sodium Dihydrate;Gluconolactone; Glucuronic Acid; Glutamic Acid, Dl-; Glutathione;Glycerin; Glycerol Ester Of Hydrogenated Rosin; Glyceryl Citrate;Glyceryl Isostearate; Glyceryl Laurate; Glyceryl Monostearate; GlycerylOleate; Glyceryl Oleate/Propylene Glycol; Glyceryl Palmitate; GlycerylRicinoleate; Glyceryl Stearate; Glyceryl Stearate-Laureth-23; GlycerylStearate/Peg Stearate; Glyceryl Stearate/Peg-100 Stearate; GlycerylStearate/Peg-40 Stearate; Glyceryl Stearate-StearamidoethylDiethylamine; Glyceryl Trioleate; Glycine; Glycine Hydrochloride; GlycolDistearate; Glycol Stearate; Guanidine Hydrochloride; Guar Gum; HairConditioner (18n195-1m); Heptane; Hetastarch; Hexylene Glycol; HighDensity Polyethylene; Histidine; Human Albumin Microspheres; HyaluronateSodium; Hydrocarbon; Hydrocarbon Gel, Plasticized; Hydrochloric Acid;Hydrochloric Acid, Diluted; Hydrocortisone; Hydrogel Polymer; HydrogenPeroxide; Hydrogenated Castor Oil; Hydrogenated Palm Oil; HydrogenatedPalm/Palm Kernel Oil Peg-6 Esters; Hydrogenated Polybutene 635-690;Hydroxide Ion; Hydroxyethyl Cellulose; Hydroxyethylpiperazine EthaneSulfonic Acid; Hydroxymethyl Cellulose; HydroxyoctacosanylHydroxystearate; Hydroxypropyl Cellulose; Hydroxypropyl Methylcellulose2906; Hydroxypropyl-Beta-cyclodextrin; Hypromellose 2208 (15000 Mpa.S);Hypromellose 2910 (15000 Mpa.S); Hypromelloses; Imidurea; Iodine;Iodoxamic Acid; Iofetamine Hydrochloride; Irish Moss Extract; Isobutane;Isoceteth-20; Isoleucine; Isooctyl Acrylate; Isopropyl Alcohol;Isopropyl Isostearate; Isopropyl Myristate; Isopropyl Myristate-MyristylAlcohol; Isopropyl Palmitate; Isopropyl Stearate; Isostearic Acid;Isostearyl Alcohol; Isotonic Sodium Chloride Solution; Jelene; Kaolin;Kathon Cg; Kathon Cg II; Lactate; Lactic Acid; Lactic Acid, Dl-; LacticAcid, L-; Lactobionic Acid; Lactose; Lactose Monohydrate; Lactose,Hydrous; Laneth; Lanolin; Lanolin Alcohol-Mineral Oil; Lanolin Alcohols;Lanolin Anhydrous; Lanolin Cholesterols; Lanolin Nonionic Derivatives;Lanolin, Ethoxylated; Lanolin, Hydrogenated; Lauralkonium Chloride;Lauramine Oxide; Laurdimonium Hydrolyzed Animal Collagen; LaurethSulfate; Laureth-2; Laureth-23; Laureth-4; Lauric Diethanolamide; LauricMyristic Diethanolamide; Lauroyl Sarcosine; Lauryl Lactate; LaurylSulfate; Lavandula Angustifolia Flowering Top; Lecithin; LecithinUnbleached; Lecithin, Egg; Lecithin, Hydrogenated; Lecithin,Hydrogenated Soy; Lecithin, Soybean; Lemon Oil; Leucine; Levulinic Acid;Lidofenin; Light Mineral Oil; Light Mineral Oil (85 Ssu); Limonene,(+/−)-; Lipocol Sc-15; Lysine; Lysine Acetate; Lysine Monohydrate;Magnesium Aluminum Silicate; Magnesium Aluminum Silicate Hydrate;Magnesium Chloride; Magnesium Nitrate; Magnesium Stearate; Maleic Acid;Mannitol; Maprofix; Mebrofenin; Medical Adhesive Modified S-15; MedicalAntiform A-F Emulsion; Medronate Disodium; Medronic Acid; Meglumine;Menthol; Metacresol; Metaphosphoric Acid; Methanesulfonic Acid;Methionine; Methyl Alcohol; Methyl Gluceth-10; Methyl Gluceth-20; MethylGluceth-20 Sesquistearate; Methyl Glucose Sesquistearate; MethylLaurate; Methyl Pyrrolidone; Methyl Salicylate; Methyl Stearate;Methylboronic Acid; Methylcellulose (4000 Mpa. S); Methylcelluloses;Methylchloroisothiazolinone; Methylene Blue; Methylisothiazolinone;Methylparaben; Microcrystalline Wax; Mineral Oil; Mono And Diglyceride;Monostearyl Citrate; Monothioglycerol; Multisterol Extract; MyristylAlcohol; Myristyl Lactate; Myristyl-.Gamma.-Picolinium Chloride;N-(Carbamoyl-Methoxy Peg-40)-1,2-Distearoyl-Cephalin Sodium;N,N-Dimethylacetamide; Niacinamide; Nioxime; Nitric Acid; Nitrogen;Nonoxynol Iodine; Nonoxynol-15; Nonoxynol-9; Norflurane; Oatmeal;Octadecene-1/Maleic Acid Copolymer; Octanoic Acid; Octisalate;Octoxynol-1; Octoxynol-40; Octoxynol-9; Octyldodecanol; OctylphenolPolymethylene; Oleic Acid; Oleth-10/Oleth-5; Oleth-2; Oleth-20; OleylAlcohol; Oleyl Oleate; Olive Oil; Oxidronate Disodium; Oxyquinoline;Palm Kernel Oil; Palmitamine Oxide; Parabens; Paraffin; Paraffin, WhiteSoft; Parfum Creme 45/3; Peanut Oil; Peanut Oil, Refined; Pectin; Peg6-32 Stearate/Glycol Stearate; Peg Vegetable Oil; Peg-100 Stearate;Peg-12 Glyceryl Laurate; Peg-120 Glyceryl Stearate; Peg-120 MethylGlucose Dioleate; Peg-15 Cocamine; Peg-150 Distearate; Peg-2 Stearate;Peg-20 Sorbitan Isostearate; Peg-22 Methyl Ether/Dodecyl GlycolCopolymer; Peg-25 Propylene Glycol Stearate; Peg-4 Dilaurate; Peg-4Laurate; Peg-40 Castor Oil; Peg-40 Sorbitan Diisostearate;Peg-45/Dodecyl Glycol Copolymer; Peg-5 Oleate; Peg-50 Stearate; Peg-54Hydrogenated Castor Oil; Peg-6 Isostearate; Peg-60 Castor Oil; Peg-60Hydrogenated Castor Oil; Peg-7 Methyl Ether; Peg-75 Lanolin; Peg-8Laurate; Peg-8 Stearate; Pegoxol 7 Stearate; Pentadecalactone;Pentaerythritol Cocoate; Pentasodium Pentetate; Pentetate CalciumTrisodium; Pentetic Acid; Peppermint Oil; Perflutren; Perfume 25677;Perfume Bouquet; Perfume E-1991; Perfume Gd 5604; Perfume Tana 90/42Scba; Perfume W-1952-1; Petrolatum; Petrolatum, White; PetroleumDistillates; Phenol; Phenol, Liquefied; Phenonip; Phenoxyethanol;Phenylalanine; Phenylethyl Alcohol; Phenylmercuric Acetate;Phenylmercuric Nitrate; Phosphatidyl Glycerol, Egg; Phospholipid;Phospholipid, Egg; Phospholipon 90g; Phosphoric Acid; Pine Needle Oil(Pinus Sylvestris); Piperazine Hexahydrate; Plastibase-50w; Polacrilin;Polidronium Chloride; Poloxamer 124; Poloxamer 181; Poloxamer 182;Poloxamer 188; Poloxamer 237; Poloxamer 407;Poly(Bis(P-Carboxyphenoxy)Propane Anhydride): Sebacic Acid;Poly(Dimethylsiloxane/Methylvinylsiloxane/Methylhydrogensiloxane)Dimethylvinyl Or Dimethylhydroxy Or Trimethyl Endblocked;Poly(Dl-Lactic-Co-Glycolic Acid), (50:50; Poly(Dl-Lactic-Co-GlycolicAcid), Ethyl Ester Terminated, (50:50; Polyacrylic Acid (250000 Mw);Polybutene (1400 Mw); Polycarbophil; Polyester; Polyester PolyamineCopolymer; Polyester Rayon; Polyethylene Glycol 1000; PolyethyleneGlycol 1450; Polyethylene Glycol 1500; Polyethylene Glycol 1540;Polyethylene Glycol 200; Polyethylene Glycol 300; Polyethylene Glycol300-1600; Polyethylene Glycol 3350; Polyethylene Glycol 400;Polyethylene Glycol 4000; Polyethylene Glycol 540; Polyethylene Glycol600; Polyethylene Glycol 6000; Polyethylene Glycol 8000; PolyethyleneGlycol 900; Polyethylene High Density Containing Ferric Oxide Black(<1%); Polyethylene Low Density Containing Barium Sulfate (20-24%);Polyethylene T; Polyethylene Terephthalates; Polyglactin; Polyglyceryl-3Oleate; Polyglyceryl-4 Oleate; Polyhydroxyethyl Methacrylate;Polyisobutylene; Polyisobutylene (1100000 Mw); Polyisobutylene (35000Mw); Polyisobutylene 178-236; Polyisobutylene 241-294; Polyisobutylene35-39; Polyisobutylene Low Molecular Weight; Polyisobutylene MediumMolecular Weight; Polyisobutylene/Polybutene Adhesive; Polylactide;Polyols; Polyoxyethylene-Polyoxypropylene 1800; PolyoxyethyleneAlcohols; Polyoxyethylene Fatty Acid Esters; Polyoxyethylene Propylene;Polyoxyl 20 Cetostearyl Ether; Polyoxyl 35 Castor Oil; Polyoxyl 40Hydrogenated Castor Oil; Polyoxyl 40 Stearate; Polyoxyl 400 Stearate;Polyoxyl 6 And Polyoxyl 32 Palmitostearate; Polyoxyl Distearate;Polyoxyl Glyceryl Stearate; Polyoxyl Lanolin; Polyoxyl Palmitate;Polyoxyl Stearate; Polypropylene; Polypropylene Glycol;Polyquaternium-10; Polyquaternium-7 (70/30 Acrylamide/Dadmac;Polysiloxane; Polysorbate 20; Polysorbate 40; Polysorbate 60;Polysorbate 65; Polysorbate 80; Polyurethane; Polyvinyl Acetate;Polyvinyl Alcohol; Polyvinyl Chloride; Polyvinyl Chloride-PolyvinylAcetate Copolymer; Polyvinylpyridine; Poppy Seed Oil; Potash; PotassiumAcetate; Potassium Alum; Potassium Bicarbonate; Potassium Bisulfite;Potassium Chloride; Potassium Citrate; Potassium Hydroxide; PotassiumMetabisulfite; Potassium Phosphate, Dibasic; Potassium Phosphate,Monobasic; Potassium Soap; Potassium Sorbate; Povidone AcrylateCopolymer; Povidone Hydrogel; Povidone K17; Povidone K25; PovidoneK29/32; Povidone K30; Povidone K90; Povidone K90f; Povidone/EicoseneCopolymer; Povidones; Ppg-12/Smdi Copolymer; Ppg-15 Stearyl Ether;Ppg-20 Methyl Glucose Ether Distearate; Ppg-26 Oleate; Product Wat;Proline; Promulgen D; Promulgen G; Propane; Propellant A-46; PropylGallate; Propylene Carbonate; Propylene Glycol; Propylene GlycolDiacetate; Propylene Glycol Dicaprylate; Propylene Glycol Monolaurate;Propylene Glycol Monopalmitostearate; Propylene Glycol Palmitostearate;Propylene Glycol Ricinoleate; Propylene Glycol/DiazolidinylUrea/Methylparaben/Propylparben; Propylparaben; Protamine Sulfate;Protein Hydrolysate; Pvm/Ma Copolymer; Quaternium-15; Quaternium-15Cis-Form; Quaternium-52; Ra-2397; Ra-3011; Saccharin; Saccharin Sodium;Saccharin Sodium Anhydrous; Safflower Oil; Sd Alcohol 3a; Sd Alcohol 40;Sd Alcohol 40-2; Sd Alcohol 40b; Sepineo P 600; Serine; Sesame Oil; SheaButter; Silastic Brand Medical Grade Tubing; Silastic Medical Adhesive,Silicone Type A; Silica, Dental; Silicon; Silicon Dioxide; SiliconDioxide, Colloidal; Silicone; Silicone Adhesive 4102; Silicone Adhesive4502; Silicone Adhesive Bio-Psa Q7-4201; Silicone Adhesive Bio-PsaQ7-4301; Silicone Emulsion; Silicone/Polyester Film Strip; Simethicone;Simethicone Emulsion; Sipon Ls 20np; Soda Ash; Sodium Acetate; SodiumAcetate Anhydrous; Sodium Alkyl Sulfate; Sodium Ascorbate; SodiumBenzoate; Sodium Bicarbonate; Sodium Bisulfate; Sodium Bisulfite; SodiumBorate; Sodium Borate Decahydrate; Sodium Carbonate; Sodium CarbonateDecahydrate; Sodium Carbonate Monohydrate; Sodium Cetostearyl Sulfate;Sodium Chlorate; Sodium Chloride; Sodium Chloride Injection; SodiumChloride Injection, Bacteriostatic; Sodium Cholesteryl Sulfate; SodiumCitrate; Sodium Cocoyl Sarcosinate; Sodium Desoxycholate; SodiumDithionite; Sodium Dodecylbenzenesulfonate; Sodium FormaldehydeSulfoxylate; Sodium Gluconate; Sodium Hydroxide; Sodium Hypochlorite;Sodium Iodide; Sodium Lactate; Sodium Lactate, L-; Sodium Laureth-2Sulfate; Sodium Laureth-3 Sulfate; Sodium Laureth-5 Sulfate; SodiumLauroyl Sarcosinate; Sodium Lauryl Sulfate; Sodium Lauryl Sulfoacetate;Sodium Metabisulfite; Sodium Nitrate; Sodium Phosphate; Sodium PhosphateDihydrate; Sodium Phosphate, Dibasic; Sodium Phosphate, Dibasic,Anhydrous; Sodium Phosphate, Dibasic, Dihydrate; Sodium Phosphate,Dibasic, Dodecahydrate; Sodium Phosphate, Dibasic, Heptahydrate; SodiumPhosphate, Monobasic; Sodium Phosphate, Monobasic, Anhydrous; SodiumPhosphate, Monobasic, Dihydrate; Sodium Phosphate, Monobasic,Monohydrate; Sodium Polyacrylate (2500000 Mw); Sodium Pyrophosphate;Sodium Pyrrolidone Carboxylate; Sodium Starch Glycolate; SodiumSuccinate Hexahydrate; Sodium Sulfate; Sodium Sulfate Anhydrous; SodiumSulfate Decahydrate; Sodium Sulfite; Sodium Sulfosuccinated UndecyclenicMonoalkylolamide; Sodium Tartrate; Sodium Thioglycolate; SodiumThiomalate; Sodium Thiosulfate; Sodium Thiosulfate Anhydrous; SodiumTrimetaphosphate; Sodium Xylenesulfonate; Somay 44; Sorbic Acid;Sorbitan; Sorbitan Isostearate; Sorbitan Monolaurate; SorbitanMonooleate; Sorbitan Monopalmitate; Sorbitan Monostearate; SorbitanSesquioleate; Sorbitan Trioleate; Sorbitan Tri stearate; Sorbitol;Sorbitol Solution; Soybean Flour; Soybean Oil; Spearmint Oil;Spermaceti; Squalane; Stabilized Oxychloro Complex; Stannous2-Ethylhexanoate; Stannous Chloride; Stannous Chloride Anhydrous;Stannous Fluoride; Stannous Tartrate; Starch; Starch 1500,Pregelatinized; Starch, Corn; Stearalkonium Chloride; StearalkoniumHectorite/Propylene Carbonate; Stearamidoethyl Diethylamine;Steareth-10; Steareth-100; Steareth-2; Steareth-20; Steareth-21;Steareth-40; Stearic Acid; Stearic Diethanolamide;Stearoxytrimethylsilane; Steartrimonium Hydrolyzed Animal Collagen;Stearyl Alcohol; Sterile Water For Inhalation; Styrene/Isoprene/StyreneBlock Copolymer; Succimer; Succinic Acid; Sucralose; Sucrose; SucroseDistearate; Sucrose Polyesters; Sulfacetamide Sodium; Sulfobutylether.Beta.-Cyclodextrin; Sulfur Dioxide; Sulfuric Acid; Sulfurous Acid;Surfactol Qs; Tagatose, D-; Talc; Tall Oil; Tallow Glycerides; TartaricAcid; Tartaric Acid, Dl-; Tenox; Tenox-2; Tert-Butyl Alcohol; Tert-ButylHydroperoxide; Tert-Butylhydroquinone;Tetrakis(2-Methoxyisobutylisocyanide)Copper(I) Tetrafluoroborate;Tetrapropyl Orthosilicate; Tetrofosmin; Theophylline; Thimerosal;Threonine; Thymol; Tin; Titanium Dioxide; Tocopherol; Tocophersolan;Total parenteral nutrition, lipid emulsion; Triacetin; Tricaprylin;Trichloromonofluoromethane; Trideceth-10; Triethanolamine LaurylSulfate; Trifluoroacetic Acid; Triglycerides, Medium Chain;Trihydroxystearin; Trilaneth-4 Phosphate; Trilaureth-4 Phosphate;Trisodium Citrate Dihydrate; Trisodium Hedta; Triton 720; Triton X-200;Trolamine; Tromantadine; Tromethamine (TRIS); Tryptophan; Tyloxapol;Tyrosine; Undecylenic Acid; Union 76 Amsco-Res 6038; Urea; Valine;Vegetable Oil; Vegetable Oil Glyceride, Hydrogenated; Vegetable Oil,Hydrogenated; Versetamide; Viscarin; Viscose/Cotton; Vitamin E; Wax,Emulsifying; Wecobee Fs; White Ceresin Wax; White Wax; Xanthan Gum;Zinc; Zinc Acetate; Zinc Carbonate; Zinc Chloride; and Zinc Oxide.

Pharmaceutical composition formulations of AAV particles disclosedherein may include cations or anions. In one embodiment, theformulations include metal cations such as, but not limited to, Zn2+,Ca2+, Cu2+, Mn2+, Mg+ and combinations thereof. As a non-limitingexample, formulations may include polymers and complexes with a metalcation (See e.g., U.S. Pat. Nos. 6,265,389 and 6,555,525, each of whichis herein incorporated by reference in its entirety). Formulations ofthe disclosure may also include one or more pharmaceutically acceptablesalts.

As used herein, “pharmaceutically acceptable salts” refers toderivatives of the disclosed compounds wherein the parent compound ismodified by converting an existing acid or base moiety to its salt form(e.g., by reacting the free base group with a suitable organic acid).Examples of pharmaceutically acceptable salts include, but are notlimited to, mineral or organic acid salts of basic residues such asamines; alkali or organic salts of acidic residues such as carboxylicacids; and the like. Representative acid addition salts include acetate,acetic acid, adipate, alginate, ascorbate, aspartate, benzenesulfonate,benzene sulfonic acid, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate,glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide,hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like, aswell as nontoxic ammonium, quaternary ammonium, and amine cations,including, but not limited to ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, ethylamine, and the like. The pharmaceutically acceptablesalts of the present disclosure include the conventional non-toxic saltsof the parent compound formed, for example, from non-toxic inorganic ororganic acids.

The pharmaceutically acceptable salts of the present disclosure can besynthesized from the parent compound which contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha stoichiometric amount of the appropriate base or acid in water or inan organic solvent, or in a mixture of the two; generally, nonaqueousmedia like ether, ethyl acetate, ethanol, isopropanol, or acetonitrileare preferred. Lists of suitable salts are found in Remington'sPharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa.,1985, p. 1418, Pharmaceutical Salts: Properties, Selection, and Use, P.H. Stahl and C. G. Wermuth (eds.), Wiley-VCH, 2008, and Berge et al.,Journal of Pharmaceutical Science, 66, 1-19 (1977); the content of eachof which is incorporated herein by reference in their entirety.

The term “pharmaceutically acceptable solvate,” as used herein, means acompound of the disclosure wherein molecules of a suitable solvent areincorporated in the crystal lattice. A suitable solvent isphysiologically tolerable at the dosage administered. Solvates may beprepared by crystallization, recrystallization, or precipitation from asolution that includes organic solvents, water, or a mixture thereof.Examples of suitable solvents are ethanol, water (for example, mono-,di-, and tri-hydrates), N-methylpyrrolidinone (NMP), dimethyl sulfoxide(DMSO), N,N′-dimethylformamide (DMF), N,N′-dimethylacetamide (DMAC),1,3-dimethyl-2-imidazolidinone (DMEU),1,3-dimethyl-3,4,5,6-tetrahydro-2-(1H)-pyrimidinone (DMPU), acetonitrile(ACN), propylene glycol, ethyl acetate, benzyl alcohol, 2-pyrrolidone,benzyl benzoate, and the like. When water is the solvent, the solvate isreferred to as a “hydrate.”

III. Administration and Dosing Administration

In one embodiment, the AAV particle may be administered to a subject(e.g., to the CNS of a subject) in a therapeutically effective amount toreduce the symptoms of the disease of the central nervous system (e.g.,Parkinson's Disease) of a subject (e.g., determined using a knownevaluation method).

The AAV particles of the present disclosure may be administered by anydelivery route which results in a therapeutically effective outcome.These include, but are not limited to, enteral (into the intestine),gastroenteral, epidural (into the dura mater), oral (by way of themouth), transdermal, intracerebral (into the cerebrum),intracerebroventricular (into the cerebral ventricles), epicutaneous(application onto the skin), intradermal, (into the skin itself),subcutaneous (under the skin), nasal administration (through the nose),intravenous (into a vein), intravenous bolus, intravenous drip,intra-arterial (into an artery), intramuscular (into a muscle),intracardiac (into the heart), intraosseous infusion (into the bonemarrow), intrathecal (into the spinal canal), intraparenchymal (intobrain tissue), intraperitoneal, (infusion or injection into theperitoneum), intravesical infusion, intravitreal, (through the eye),intracavernous injection (into a pathologic cavity) intracavitary (intothe base of the penis), intravaginal administration, intrauterine,extra-amniotic administration, transdermal (diffusion through the intactskin for systemic distribution), transmucosal (diffusion through amucous membrane), transvaginal, insufflation (snorting), sublingual,sublabial, enema, eye drops (onto the conjunctiva), or in ear drops,auricular (in or by way of the ear), buccal (directed toward the cheek),conjunctival, cutaneous, dental (to a tooth or teeth), electro-osmosis,endocervical, endosinusial, endotracheal, extracorporeal, hemodialysis,infiltration, interstitial, intra-abdominal, intra-amniotic,intra-articular, intrabiliary, intrabronchial, intrabursal,intracartilaginous (within a cartilage), intracaudal (within the caudaequine), intracisternal (within the cisterna magna cerebellomedularis),intracorneal (within the cornea), dental intracoronal, intracoronary(within the coronary arteries), intracorporus cavernosum (within thedilatable spaces of the corporus cavernosa of the penis), intradiscal(within a disc), intraductal (within a duct of a gland), intraduodenal(within the duodenum), intradural (within or beneath the dura),intraepidermal (to the epidermis), intraesophageal (to the esophagus),intragastric (within the stomach), intragingival (within the gingivae),intraileal (within the distal portion of the small intestine),intralesional (within or introduced directly to a localized lesion),intraluminal (within a lumen of a tube), intralymphatic (within thelymph), intramedullary (within the marrow cavity of a bone),intrameningeal (within the meninges), intramyocardial (within themyocardium), intraocular (within the eye), intraovarian (within theovary), intrapericardial (within the pericardium), intrapleural (withinthe pleura), intraprostatic (within the prostate gland), intrapulmonary(within the lungs or its bronchi), intrasinal (within the nasal orperiorbital sinuses), intraspinal (within the vertebral column),intrasynovial (within the synovial cavity of a joint), intratendinous(within a tendon), intratesticular (within the testicle), intrathecal(within the cerebrospinal fluid at any level of the cerebrospinal axis),intrathoracic (within the thorax), intratubular (within the tubules ofan organ), intratumor (within a tumor), intratympanic (within the aurusmedia), intravascular (within a vessel or vessels), intraventricular(within a ventricle), iontophoresis (by means of electric current whereions of soluble salts migrate into the tissues of the body), irrigation(to bathe or flush open wounds or body cavities), laryngeal (directlyupon the larynx), nasogastric (through the nose and into the stomach),occlusive dressing technique (topical route administration which is thencovered by a dressing which occludes the area), ophthalmic (to theexternal eye), oropharyngeal (directly to the mouth and pharynx),parenteral, percutaneous, periarticular, peridural, perineural,periodontal, rectal, respiratory (within the respiratory tract byinhaling orally or nasally for local or systemic effect), retrobulbar(behind the pons or behind the eyeball), soft tissue, subarachnoid,subconjunctival, submucosal, topical, transplacental (through or acrossthe placenta), transtracheal (through the wall of the trachea),transtympanic (across or through the tympanic cavity), ureteral (to theureter), urethral (to the urethra), vaginal, caudal block, diagnostic,nerve block, biliary perfusion, cardiac perfusion, photopheresis andspinal.

In some embodiments, compositions may be administered in a way whichallows them to cross the blood-brain barrier, vascular barrier, or otherepithelial barrier. The AAV particles of the present disclosure may beadministered in any suitable form, either as a liquid solution orsuspension, as a solid form suitable for liquid solution or suspensionin a liquid solution. The AAV particles may be formulated with anyappropriate and pharmaceutically acceptable excipient.

In one embodiment, the AAV particles of the present disclosure may bedelivered to a subject via a single route administration.

In one embodiment, the AAV particles of the present disclosure may bedelivered to a subject via a multi-site route of administration. Asubject may be administered at 2, 3, 4, 5 or more than 5 sites.

In one embodiment, a subject may be administered the AAV particles ofthe present disclosure using a bolus infusion.

In one embodiment, a subject may be administered the AAV particles ofthe present disclosure using sustained delivery over a period ofminutes, hours or days. The infusion rate may be changed depending onthe subject, distribution, formulation or another delivery parameter.

In one embodiment, the AAV particles of the present disclosure may bedelivered by intramuscular delivery route. (See, e.g., U.S. Pat. No.6,506,379; the content of which is incorporated herein by reference inits entirety). Non-limiting examples of intramuscular administrationinclude an intravenous injection or a subcutaneous injection.

In one embodiment, the AAV particles of the present disclosure may bedelivered by oral administration. Non-limiting examples of oraladministration include a digestive tract administration and a buccaladministration.

In one embodiment, the AAV particles of the present disclosure may bedelivered by intraocular delivery route. A non-limiting example ofintraocular administration include an intravitreal injection.

In one embodiment, the AAV particles of the present disclosure may bedelivered by intranasal delivery route. Non-limiting examples ofintranasal delivery include administration of nasal drops or nasalsprays.

In some embodiments, the AAV particles that may be administered to asubject by peripheral injections. Non-limiting examples of peripheralinjections include intraperitoneal, intramuscular, intravenous,conjunctival or joint injection. It was disclosed in the art that theperipheral administration of AAV particles can be transported to thecentral nervous system, for example, to the motor neurons (e.g., U. S.Patent Publication Nos. 20100240739; and 20100130594; the content ofeach of which is incorporated herein by reference in their entirety).

In one embodiment, the AAV particles may be delivered by injection intothe CSF pathway. Non-limiting examples of delivery to the CSF pathwayinclude intrathecal and intracerebroventricular administration.

In one embodiment, the AAV particles may be delivered by systemicdelivery. As a non-limiting example, the systemic delivery may be byintravascular administration.

In one embodiment, the AAV particles of the present disclosure may beadministered to a subject by intracranial delivery (See, e.g., U.S. Pat.No. 8,119,611; the content of which is incorporated herein by referencein its entirety).

In some embodiments, the AAV particles of the present disclosure may beadministered by injection. As a non-limiting example, the AAV particlesof the present disclosure may be administered to a subject by injection.

In some embodiments, the AAV particles of the present disclosure may beadministered by muscular injection. As a non-limiting example, the AAVparticles of the present disclosure may be administered to a subject bymuscular administration.

In some embodiments, the AAV particles of the present disclosure may beadministered by intramuscular administration. As a non-limiting example,the AAV particles of the present disclosure may be administered to asubject by intramuscular administration.

In one embodiment, the AAV particles of the present disclosure areadministered to a subject and transduce muscle of a subject. As anon-limiting example, the AAV particles are administered byintramuscular administration.

In some embodiments, the AAV particles of the present disclosure may beadministered via intraparenchymal injection. As a non-limiting example,the AAV particles of the present disclosure may be administered to asubject by intraparenchymal administration.

In some embodiments, the AAV particles of the present disclosure may beadministered by intravenous administration. As a non-limiting example,the AAV particles of the present disclosure may be administered to asubject by intravenous administration.

In one embodiment, the AAV particles of the present disclosure may beadministered via intravenous delivery.

In one embodiment, the AAV particles of the present disclosure may beadministered via a single dose intravenous delivery. As a non-limitingexample, the single dose intravenous delivery may be a one-timetreatment. In the context of diseases of the central nervous system(e.g., Parkinson's Disease), the single dose intravenous delivery canproduce durable relief for subjects with central nervous system (e.g.,Parkinson's Disease) and/or related symptoms. The relief may last forminutes such as, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 minutes or more than 59minutes; hours such as, but not limited to, 1, 2, 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, or more than 48 hours; days such as, but not limitedto, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or more than 31 days;weeks such as, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, or more than 16 weeks; months such as, but notlimited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, or more than 24 months; years such as, butnot limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, ormore than 15 years.

In one embodiment, the AAV particles of the present disclosure may beadministered via intravenous delivery to the DRG nociceptive neurons.

In one embodiment, the AAV particles of the present disclosure may beadministered via a single dose intravenous delivery to the DRGnociceptive neurons. As a non-limiting example, the single doseintravenous delivery may be a one-time treatment. In the context ofdiseases of the central nervous system (e.g., Parkinson's Disease), thesingle dose intravenous delivery can produce durable relief for subjectswith diseases of the central nervous system (e.g., Parkinson's Disease)and/or related symptoms. The relief may last for minutes such as, butnot limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34,35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52,53, 54, 55, 56, 57, 58, 59 minutes or more than 59 minutes; hours suchas, but not limited to, 1, 2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, ormore than 48 hours; days such as, but not limited to, 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, or more than 31 days; weeks such as, but notlimited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, ormore than 16 weeks; months such as, but not limited to, 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,or more than 24 months; years such as, but not limited to, 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or more than 15 years.

In some embodiments, the AAV particles of the present disclosure may beadministered by intrathecal injection. As a non-limiting example, theAAV particles of the present disclosure may be administered byintrathecal injection.

In one embodiment, the AAV particle may be administered to the cisternamagna in a therapeutically effective amount to transduce spinal cordmotor neurons and/or astrocytes. As a non-limiting example, the AAVparticle may be administered intrathecally.

In one embodiment, the AAV particle may be administered usingintrathecal infusion in a therapeutically effective amount to transducespinal cord motor neurons and/or astrocytes.

In some embodiments, the AAV particles of the present disclosure may beadministered via a single dose intrathecal injection. As a non-limitingexample, the single dose intrathecal injection may be a one-timetreatment. In the context of diseases of the central nervous system(e.g., Parkinson's Disease), the single dose intrathecal injection canproduce durable relief for subjects with diseases of the central nervoussystem (e.g., Parkinson's Disease) and/or related symptoms. The reliefmay last for minutes such as, but not limited to, 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 minutesor more than 59 minutes; hours such as, but not limited to, 1, 2, 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48, or more than 48 hours; days such as, butnot limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or more than31 days; weeks such as, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, or more than 16 weeks; months such as, butnot limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, or more than 24 months; years such as,but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,or more than 15 years.

In some embodiments, the AAV particles of the present disclosure may beadministered via intrathecal injection to the DRG nociceptive neurons.

In some embodiments, the AAV particles of the present disclosure may beadministered via a single dose intrathecal injection to the DRGnociceptive neurons. As a non-limiting example, the single doseintrathecal injection may be a one-time treatment. In the context ofdiseases of the central nervous system (e.g., Parkinson's Disease), thesingle dose intrathecal injection can produce durable relief forsubjects with diseases of the central nervous system (e.g., Parkinson'sDisease) and/or related symptoms. The relief may last for minutes suchas, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50,51, 52, 53, 54, 55, 56, 57, 58, 59 minutes or more than 59 minutes;hours such as, but not limited to, 1, 2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,47, 48, or more than 48 hours; days such as, but not limited to, 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, or more than 31 days; weeks such as,but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, or more than 16 weeks; months such as, but not limited to, 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, or more than 24 months; years such as, but not limited to, 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or more than 15 years.

In one embodiment, the AAV particle described herein is administered viaintrathecal (IT) infusion at C1. The infusion may be for 1, 2, 3, 4, 6,7, 8, 9, 10, 11, 12, 13, 14, 15 or more than 15 hours.

In some embodiments, the AAV particles of the present disclosure may beadministered by intraparenchymal injection. As a non-limiting example,the AAV particles of the present disclosure may be administered to asubject by intraparenchymal injection.

In one embodiment, the AAV particle may be administered to the cisternamagna in a therapeutically effective amount to transduce spinal cordmotor neurons and/or astrocytes. As a non-limiting example, the AAVparticle may be administered intraparenchymal injection.

In some embodiments, the AAV particles of the present disclosure may beadministered by intraparenchymal injection and intrathecal injection. Asa non-limiting example, the AAV particles of the present disclosure maybe administered via intraparenchymal injection and intrathecalinjection.

In some embodiments, the AAV particles of the present disclosure may beadministered by subcutaneous injection. As a non-limiting example, theAAV particles In one embodiment, the AAV particles of the presentdisclosure may be administered to a subject by subcutaneous injection.

In some embodiments, the AAV particles of the present disclosure may beadministered topically. As a non-limiting example, the AAV particles ofthe present disclosure may be administered to a subject topically.

In one embodiment, the AAV particles may be delivered by directinjection into the brain. As a non-limiting example, the brain deliverymay be by intrastriatal administration.

In one embodiment, the AAV particles of the present disclosure may beadministered via intrastriatal injection.

In one embodiment, the AAV particles of the present disclosure may beadministered via intrastriatal injection and another route ofadministration described herein.

In one embodiment, the AAV particles may be delivered by more than oneroute of administration. As non-limiting examples of combinationadministrations, AAV particles may be delivered by intrathecal andintracerebroventricular, or by intravenous and intraparenchymaladministration.

In one embodiment, the AAV particle may be administered to the CNS in atherapeutically effective amount to improve function and/or survival fora subject with diseases of the central nervous system (e.g., Parkinson'sDisease). As a non-limiting example, the vector may be administered bydirect infusion into the striatum.

The AAV particle may be administered in a “therapeutically effective”amount, i.e., an amount that is sufficient to alleviate and/or preventat least one symptom associated with the disease, or provide improvementin the condition of the subject.

In one embodiment, the catheter may be located at more than one site inthe spine for multi-site delivery. The AAV particle may be delivered ina continuous and/or bolus infusion. Each site of delivery may be adifferent dosing regimen or the same dosing regimen may be used for eachsite of delivery. As a non-limiting example, the sites of delivery maybe in the cervical and the lumbar region. As another non-limitingexample, the sites of delivery may be in the cervical region. As anothernon-limiting example, the sites of delivery may be in the lumbar region.

In one embodiment, a subject may be analyzed for spinal anatomy andpathology prior to delivery of the AAV particle described herein. As anon-limiting example, a subject with scoliosis may have a differentdosing regimen and/or catheter location compared to a subject withoutscoliosis.

In one embodiment, the orientation of the spine of the subject duringdelivery of the AAV particle may be vertical to the ground.

In another embodiment, the orientation of the spine of the subjectduring delivery of the AAV particle may be horizontal to the ground.

In one embodiment, the spine of the subject may be at an angle ascompared to the ground during the delivery of the AAV particle. Theangle of the spine of the subject as compared to the ground may be atleast 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150or 180 degrees.

In one embodiment, the delivery method and duration is chosen to providebroad transduction in the spinal cord. As a non-limiting example,intrathecal delivery is used to provide broad transduction along therostral-caudal length of the spinal cord. As another non-limitingexample, multi-site infusions provide a more uniform transduction alongthe rostral-caudal length of the spinal cord. As yet anothernon-limiting example, prolonged infusions provide a more uniformtransduction along the rostral-caudal length of the spinal cord.

In one embodiment, administration occurs by a posterior (e.g., back ofthe head) surgical delivery approach to the putamen. As a non-limitingexample, the average putaminal coverage is 50% with posterior deliveryand the surgical time is less than 10 hours.

In one embodiment, administration occurs by a transfrontal (e.g., top ofthe head) surgical delivery approach to the putamen. As a non-limitingexample, the average putaminal coverage less than 50% with posteriordelivery and the surgical time is more than 10 hours.

Parenteral and Injectable Administration

In some embodiments, pharmaceutical compositions, AAV particles of thepresent disclosure may be administered parenterally. Liquid dosage formsfor oral and parenteral administration include, but are not limited to,pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups, and/or elixirs. In addition to active ingredients,liquid dosage forms may comprise inert diluents commonly used in the artsuch as, for example, water or other solvents, solubilizing agents andemulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate,ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol,1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed,groundnut, corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, oralcompositions can include adjuvants such as wetting agents, emulsifyingand suspending agents, sweetening, flavoring, and/or perfuming agents.In certain embodiments for parenteral administration, compositions aremixed with solubilizing agents such as CREMOPHOR®, alcohols, oils,modified oils, glycols, polysorbates, cyclodextrins, polymers, and/orcombinations thereof. In other embodiments, surfactants are includedsuch as hydroxypropylcellulose.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing agents, wetting agents, and/or suspendingagents. Sterile injectable preparations may be sterile injectablesolutions, suspensions, and/or emulsions in nontoxic parenterallyacceptable diluents and/or solvents, for example, as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution, U.S.P., and isotonic sodiumchloride solution. Sterile, fixed oils are conventionally employed as asolvent or suspending medium. For this purpose, any bland fixed oil canbe employed including synthetic mono- or diglycerides. Fatty acids suchas oleic acid can be used in the preparation of injectables.

Injectable formulations may be sterilized, for example, by filtrationthrough a bacterial-retaining filter, and/or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of active ingredients, it is oftendesirable to slow the absorption of active ingredients from subcutaneousor intramuscular injections. This may be accomplished by the use ofliquid suspensions of crystalline or amorphous material with poor watersolubility. The rate of absorption of active ingredients depends uponthe rate of dissolution which, in turn, may depend upon crystal size andcrystalline form. Alternatively, delayed absorption of a parenterallyadministered drug form is accomplished by dissolving or suspending thedrug in an oil vehicle. Injectable depot forms are made by formingmicroencapsule matrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissues.

Depot Administration

As described herein, in some embodiments, pharmaceutical compositions,AAV particles of the present disclosure are formulated in depots forextended release. Generally, specific organs or tissues (“targettissues”) are targeted for administration.

In some aspects of the disclosure, pharmaceutical compositions, AAVparticles of the present disclosure are spatially retained within orproximal to target tissues. Provided are methods of providingpharmaceutical compositions, AAV particles, to target tissues ofmammalian subjects by contacting target tissues (which comprise one ormore target cells) with pharmaceutical compositions, AAV particles,under conditions such that they are substantially retained in targettissues, meaning that least 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95,96, 97, 98, 99, 99.9, 99.99 or greater than 99.99% of the composition isretained in the target tissues. Advantageously, retention is determinedby measuring the amount of pharmaceutical compositions, AAV particles,that enter one or more target cells. For example, at least 1%, 5%, 10%,20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%,99.9%, 99.99% or greater than 99.99% of pharmaceutical compositions, AAVparticles, administered to subjects are present intracellularly at aperiod of time following administration. For example, intramuscularinjection to mammalian subjects may be performed using aqueouscompositions comprising pharmaceutical compositions, AAV particles ofthe present disclosure and one or more transfection reagents, andretention is determined by measuring the amount of pharmaceuticalcompositions, AAV particles, present in muscle cells. Certain aspects ofthe disclosure are directed to methods of providing pharmaceuticalcompositions, AAV particles of the present disclosure to a targettissues of mammalian subjects, by contacting target tissues (comprisingone or more target cells) with pharmaceutical compositions, AAVparticles under conditions such that they are substantially retained insuch target tissues.

Pharmaceutical compositions, AAV particles comprise enough activeingredient such that the effect of interest is produced in at least onetarget cell. In some embodiments, pharmaceutical compositions, AAVparticles generally comprise one or more cell penetration agents,although “naked” formulations (such as without cell penetration agentsor other agents) are also contemplated, with or without pharmaceuticallyacceptable carriers.

Delivery to the Central Nervous System

In one embodiment, delivery of the pharmaceutical compositionscomprising AAV particles to cells of the central nervous system (e.g.,parenchyma) comprises infusion of up to 1 mL. In one embodiment,delivery of the pharmaceutical compositions comprising AAV particles tocells of the central nervous system (e.g., parenchyma) may compriseinfusion of 0.001, 0.002, 0.003, 0.004, 0.005, 0.010, 0.015, 0.020,0.025, 0.030, 0.040, 0.050, 0.060, 0.070, 0.080, 0.090, 0.1, 0.2, 0.3,0.4, 0.5, 0.6, 0.7, 0.8, or 0.9 mL.

In one embodiment, delivery of pharmaceutical composition comprising AAVparticles to cells of the central nervous system (e.g., parenchyma)comprises infusion of between about 1 mL to about 120 mL. In oneembodiment, delivery of pharmaceutical composition comprising AAVparticles to cells of the central nervous system (e.g., parenchyma) maycomprise infusion of 0.1, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8,1.9, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38,39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56,57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92,93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108,109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, or 120 mL. In oneembodiment, delivery of AAV particles to cells of the central nervoussystem (e.g., parenchyma) comprises infusion of at least 3 mL. In oneembodiment, delivery of AAV particles to cells of the central nervoussystem (e.g., parenchyma) consists of infusion of 3 mL. In oneembodiment, delivery of AAV particles to cells of the central nervoussystem (e.g., parenchyma) comprises infusion of at least 10 mL. In oneembodiment, delivery of AAV particles to cells of the central nervoussystem (e.g., parenchyma) consists of infusion of 10 mL.

In one embodiment, the volume of the pharmaceutical compositioncomprising AAV particles delivered to the cells of the central nervoussystem (e.g., parenchyma) of a subject is 50 ul, 100 ul, 200 ul, 300 ul,400 ul, 500 ul, 600 ul, 700 ul, 800 ul, 900 ul, 1000 ul, 1100 ul, 1200ul, 1300 ul, 1400 ul, 1500 ul, 1600 ul, 1700 ul, 1800 ul, 1900 ul, 2000ul or more than 2000 ul.

In one embodiment, the volume of the pharmaceutical compositioncomprising AAV particles delivered to a region in both hemispheres of asubject brain is 50 ul, 100 ul, 200 ul, 300 ul, 400 ul, 500 ul, 600 ul,700 ul, 800 ul, 900 ul, 1000 ul, 1100 ul, 1200 ul, 1300 ul, 1400 ul,1500 ul, 1600 ul, 1700 ul, 1800 ul, 1900 ul, 2000 ul or more than 2000ul. As a non-limiting example, the volume delivered to a region in bothhemispheres is 200 ul. As another non-limiting example, the volumedelivered to a region in both hemispheres is 900 ul. As yet anothernon-limiting example, the volume delivered to a region in bothhemispheres is 1800 ul.

In one embodiment, the volume of the pharmaceutical compositioncomprising AAV particles delivered to the putamen in both hemispheres ofa subject brain is 50 ul, 100 ul, 200 ul, 300 ul, 400 ul, 450 ul, 500ul, 600 ul, 700 ul, 800 ul, 900 ul, 1000 ul, 1100 ul, 1200 ul, 1300 ul,1400 ul, 1500 ul, 1600 ul, 1700 ul, 1800 ul, 1900 ul, 2000 ul or morethan 2000 ul. As a non-limiting example, the volume delivered to theputamen in both hemispheres is 100 ul. As another non-limiting example,the volume delivered to the putamen in both hemispheres is 200 ul. As anon-limiting example, the volume delivered to the putamen in bothhemispheres is 300 ul. As another non-limiting example, the volumedelivered to the putamen in both hemispheres is 450 ul. As anothernon-limiting example, the volume delivered to the putamen in bothhemispheres is 900 ul. As yet another non-limiting example, the volumedelivered to the putamen both hemispheres is 1800 ul.

In one embodiment, the volume of the pharmaceutical compositioncomprising AAV particles delivered to a subject is 900 ul to eachputamen.

In one embodiment, the volume of the pharmaceutical compositioncomprising AAV particles delivered to a subject is 450 ul to eachputamen.

In one embodiment, the total volume delivered to a subject may be splitbetween one or more administration sites e.g., 1, 2, 3, 4, 5 or morethan 5 sites. As a non-limiting example, the total volume is splitbetween administration to the left and right putamen. As anothernon-limiting example, the total volume is split between two sites ofadministration to each of the left and right putamen.

In one embodiment, the pharmaceutical composition comprising AAVparticles is administered using a fenestrated needle. Non-limitingexamples of fenestrated needles are described in U.S. Pat. Nos.8,333,734, 7,135,010, 7,575,572, 7,699,852, 4,411,657, 6,890,319,6,613,026, 6,726,659, 6,565,572, 6,520,949, 6,382,212, 5,848,996,5,759,179, 5,674,267, 5,588,960, 5,484,401, 5,199,441, 5,012,818,4,474,569, 3,766,907, 3,552,394, the contents of each of which areherein incorporated by reference in its entirety.

In one embodiment, a composition comprises at least one payloaddescribed herein and the payloads are components of a viral genomepackaged in an AAV particle. The percent (%) ratio of AAV particlescomprising the payload to the AAV particles without the payload (alsoreferred to herein as empty capsids) in the composition may be 0:100,1:99, 0: 90, 15:85, 25:75, 30:70, 50:50, 70:30, 85:15, 90:10, 99:1 or100:0. As a non-limiting example, the percent ratio of AAV particlescomprising the payload to empty capsids is 50:50. As anothernon-limiting example, the percent ratio of AAV particles comprising thepayload to empty capsids is 70:30. As another non-limiting example, thepercent ratio of AAV particles comprising the payload to empty capsidsis 85:15. As another non-limiting example, the percent ratio of AAVparticles comprising the payload to empty capsids is 100:0.

In one embodiment, the composition described herein comprises at least1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 51, 52, 53, 54, 55, 60, 65,70, 75, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,96, 97, 98, 99 or greater than 99% AAV particles comprising the payload.As a non-limiting example, the composition comprises at least 50% AAVparticles comprising the payload. As another non-limiting example, thecomposition comprises at least 52% AAV particles comprising the payload.As another non-limiting example, the composition comprises at least 58%AAV particles comprising the payload. As another non-limiting example,the composition comprises at least 70% AAV particles comprising thepayload. As another non-limiting example, the composition comprises atleast 83% AAV particles comprising the payload. As another non-limitingexample, the composition comprises at least 85% AAV particles comprisingthe payload. As another non-limiting example, the composition comprisesat least 99% AAV particles comprising the payload. As anothernon-limiting example, the composition comprises 100% AAV particlescomprising the payload.

In one embodiment, the composition described herein comprises 1-10%,10-20%, 30-40%, 50-60%, 50-70%, 50-80%, 50-90%, 50-99%, 50-100%, 60-70%,60-80%, 60-90%, 60-99%, 60-100%, 70-80%, 70-90%, 70-99%, 70-100%,80-85%, 80-90%, 80-95%, 80-99%, 80-100%, 90-95%, 90-99%, or 90-100% AAVparticles comprising the payload. As a non-limiting example, thecomposition described herein comprises 50-100% AAV particles comprisingthe payload. As another non-limiting example, the composition describedherein comprises 50-60% AAV particles comprising the payload. As anothernon-limiting example, the composition described herein comprises 80-99%AAV particles comprising the payload. As another non-limiting example,the composition described herein comprises 80-90% AAV particlescomprising the payload. As a non-limiting example, the compositiondescribed herein comprises 80-95% AAV particles comprising the payload.As a non-limiting example, the composition described herein comprises80-85% AAV particles comprising the payload.

In one embodiment, the composition described herein comprises less than1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 51, 52, 53, 54, 55, 60, 65,70, 75, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,96, 97, 98, 99 or 100% empty particles. As a non-limiting example, thecomposition comprises less than 50% empty particles. As a non-limitingexample, the composition comprises less than 45% empty particles. As anon-limiting example, the composition comprises less than 40% emptyparticles. As a non-limiting example, the composition comprises lessthan 35% empty particles. As a non-limiting example, the compositioncomprises less than 30% empty particles. As a non-limiting example, thecomposition comprises less than 25% empty particles. As a non-limitingexample, the composition comprises less than 20% empty particles. As anon-limiting example, the composition comprises less than 15% emptyparticles. As a non-limiting example, the composition comprises lessthan 10% empty particles. As a non-limiting example, the compositioncomprises less than 5% empty particles. As a non-limiting example, thecomposition comprises less than 1% empty particles.

In the composition described herein comprises 1-10%, 10-20%, 30-40%,50-60%, 50-70%, 50-80%, 50-90%, 50-99%, 50-100%, 60-70%, 60-80%, 60-90%,60-99%, 60-100%, 70-80%, 70-90%, 70-99%, 70-100%, 80-85%, 80-90%,80-95%, 80-99%, 80-100%, 90-95%, 90-99%, or 90-100% empty particles. Asa non-limiting example, the composition described herein comprises30-40% empty particles. As another non-limiting example, the compositiondescribed herein comprises 30-50% empty particles. As anothernon-limiting example, the composition described herein comprises 30-60%empty particles. As another non-limiting example, the compositiondescribed herein comprises 30-70% empty particles. As a non-limitingexample, the composition described herein comprises 30-80% emptyparticles. As a non-limiting example, the composition described hereincomprises 30-90% empty particles.

In one embodiment, the ratio of distribution volume in the parenchyma ofan area of a subject to the infusion volume of an area of a subject maybe 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4,1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8,2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2,4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6,5.7, 5.8, 5.9, 6.0 or more than 6.0. As a non-limiting example, theratio of distribution volume in the parenchyma to infusion volume was1.6 in the caudate nucleus. As a non-limiting example, the ratio ofdistribution volume in the parenchyma to infusion volume was 3.1 in theputamen. As a non-limiting example, the distribution of the AAVparticles in the putamen may be 2-3 times the volume infused.

In one embodiment, the effectiveness of the dose, route ofadministration and/or volume of administration may be evaluated usingvarious methods described herein such as, but not limited to, PETimaging, L-DOPA challenge test (e.g., see Forsayeth et al. 2006, Mol.Ther. 14(4): 571-577), UPDRS scores and patient diaries (e.g., Hauserdiary). As a non-limiting example, a subject may have decreaseddyskinesia or periods of decreased dyskinesia after administration ofthe Pharmaceutical composition comprising AAV particles. As anothernon-limiting example, a subject may have a decrease in Parkinson'sDisease related symptoms including limited mobility and dyskinesia. Asyet another non-limiting example, a subject may show improvement in offtime and motor fluctuations. The improvement may be at least 10%, atleast 20%, at least 30%, at least 40%, at least 50%, at least 60%, atleast 70%, at least 80%, at least 90% or greater than 90%. Theimprovement may last for minutes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,15, 20, 25, 30, 35, 40, 45, 50, 55 or more than 55), hours (e.g., 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23 or more than 24), days (e.g., 1, 2, 3, 4, 5, 6 or more than 7), weeks(1, 2, 3, 4, 5, 6, 7 or more than 7), months (1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11 or more than 11) or years (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 ormore than 9).

In one embodiment, the selection of subjects for administration of theAAV particles described herein and/or the effectiveness of the dose,route of administration and/or volume of administration may be evaluatedusing imaging of the perivascular spaces (PVS) which are also known asVirchow-Robin spaces. PVS surround the arterioles and venules as theyperforate brain parenchyma and are filled with cerebrospinal fluid(CSF)/interstitial fluid. PVS are common in the midbrain, BG, andcentrum semiovale. While not wishing to be bound by theory, PVS may playa role in the normal clearance of metabolites and have been associatedwith worse cognition and several disease states including Parkinson'sdisease. PVS are usually normal in size but they can increase in size ina number of disease states. Potter et al. (Cerebrovasc Dis. 2015January; 39(4): 224-231; the contents of which are herein incorporatedby reference in its entirety) developed a grading method where theystudied a full range of PVS and rated basal ganglia, centrum semiovaleand midbrain PVS. They used the frequency and range of PVS used by Macand Lullich et al. (J Neurol Neurosurg Psychiatry. 2004 November;75(11):1519-23; the contents of which are herein incorporated byreference in its entirety) and Potter et al. gave 5 ratings to basalganglia and centrum semiovale PVS: 0 (none), 1 (1-10), 2 (11-20), 3(21-40) and 4 (>40) and 2 ratings to midbrain PVS: 0 (non visible) or 1(visible). The user guide for the rating system by Potter et al. can befound at: www.sbirc.ed.ac.uk/documents/epvs-rating-scale-user-guide.pdf.

In one embodiment, the selection of subjects for administration of theAAV particles described herein and/or the effectiveness of the dose,route of administration and/or volume of administration may be evaluatedusing positron emission tomography (PET) measurements of neuroimagingbiomarkers such as, but not limited to [¹⁸F]FDOPA. Neuroimagingbiomarkers such as [¹⁸F]FDOPA may be used to identify affectedindividuals and/or may be used to detect a nigrostriatal defect prior tothe onset of clinical manifestations. Further, PET-based criteria may beused to categorize subjects based on their nigrostriatal neuronalintegrity (e.g., abnormal, normal or uncertain nigrostriatal neuronalintegrity) (Rachette et al. Am J Med Genet B Neuropsychiatr Genet. 2006Apr. 5; 141B(3): 245-249; the contents of which are herein incorporatedby reference in its entirety).

In one embodiment, a subject who may be administered a dose of the AAVparticles described herein may have advanced PD and still respond tolevodopa therapy but the subject also experiences medically refractorymotor complications (e.g., sever motor fluctuations and/or dyskinesiasthat occur during levodopa and other dopaminergic therapies despiteadjustments in and optimization of medication). The subject may behealthy enough to undergo a neurosurgical procedure which may bedetermined by methods known in the art. As a non-limiting example, thesubject may meet the selection criteria for deep brain stimulation(DBS). The subject may have idiopathic PD, younger than 69 years of age,have pronounced responses to levodopa, have medication-refractorysymptoms (e.g., motor fluctuation and/or dyskinesia) and/or have littleor no cognitive dysfunction.

In one embodiment, a subject who may be administered a dose of the AAVparticles described herein may also suffer from dementia or cognitiveimpairment.

In one embodiment, a subject who may be administered a dose of the AAVparticles described herein may have been previously treated with thesame or similar therapeutic. In another embodiment, a subject may havebeen treated with a therapeutic which has been shown to reduce thesymptoms of Parkinson's Disease.

In one embodiment, a subject who may be administered a dose of the AAVparticles described herein may have failed to derive adequate benefitfrom standard medical therapy. As a non-limiting example, the subjectmay not have responded to treatment. As another non-limiting example, asubject may have residual disability despite treatment.

In one embodiment, a subject who may be administered a dose of the AAVparticles described herein may undergo testing to evaluate the levels ofneurotransmitter analytes to determine the effectiveness of the dose. Asa non-limiting example, CSF neurotransmitters, plasma AADC activityand/or urine VLA may be analyzed.

In one embodiment, a subject who may be administered a dose of the AAVparticles described herein may be videotaped or recorded in order tomonitor the progress of the subject during the course of treatment.

Delivery to the Putamen

In one embodiment, the AAV particles may be administered to the rightputamen and/or the left putamen. The administration may be at one ormore sites in the putamen such as, but not limited to, 2 sites, 3 sites,4 sites or more than 4 sites. As a non-limiting example, the AAVparticles are delivered to 2 sites in the left putamen and 2 sites inthe right putamen.

In one embodiment, the administration of the formulation of the AAVparticles to a subject provides coverage of the putamen of a subject(e.g., the left and/or right putamen). In one aspect, the administrationof the AAV particles may provide at least 8%, 9%, 10%, 13%, 14%, 15%,19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%,33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%,47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95% or more than 95% to the left and/or right putamen of asubject. As a non-limiting example, the coverage is at least 20%. As anon-limiting example, the coverage is at least 40%. In another aspect,the administration of the AAV particles may provide at least 8%, 9%,10%, 13%, 14%, 15%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%,29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%,43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95% or more than 95% coverage of thesurface area of the left and/or right putamen of a subject. As anon-limiting example, the total coverage is at least 20%. As anon-limiting example, the total coverage is at least 40%. In yet anotheraspect, the administration of the AAV particles may provide 10-40%,20-40%, 20-30%, 20-35%, 20-50%, 30-40%, 35-40%, 30-60%, 40-70%, 50-80%or 60-90% coverage to the left and/or right putamen of a subject or tothe total surface area of the left and/or right putamen of a subject.

In one embodiment, the administration of the formulation of the AAVparticles to a subject provides coverage of the posterior putamen of asubject (e.g., the left and/or right posterior putamen). In one aspect,the administration of the AAV particles may provide at least 10%, 15%,20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%,34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%,48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95% or more than 95% to the left and/or right posterior putamen ofa subject. As a non-limiting example, the coverage is at least 20%. As anon-limiting example, the coverage is at least 40%. In another aspect,the administration of the AAV particles may provide at least 10%, 15%,20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%,34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%,48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95% or more than 95% coverage of the surface area of the leftand/or right posterior putamen of a subject. As a non-limiting example,the total coverage is at least 20%. As a non-limiting example, the totalcoverage is at least 40%. In yet another aspect, the administration ofthe AAV particles may provide 10-40%, 20-50%, 30-60%, 40-70%, 50-80% or60-90% coverage to the left and/or right posterior putamen of a subjector to the total surface area of the left and/or right putamen of asubject.

In one embodiment, the AAV particles described herein may beadministered using acute bilateral placement of catheters into eachputamen. The placement may use magnetic resonance image (MRI)-guidedstereotactic neurosurgical techniques known in the art or describedherein. Additionally, a contrast agent such as, but not limited to agadolinium based contrast agent (e.g., PROHANCE®) may be used in theformulation to monitor and confirm the distribution of the formulation.

In one embodiment, a subject may be administered the AAV particles in abilateral stereotactic CED-assisted step infusion into the putamen(e.g., the post commissural putamen).

In one embodiment, a subject may be administered the AAV particles ofthe present disclosure at a dose of 4.5×10¹² vector genomes at a volumeof 900 ul per putamen.

In one embodiment, a subject may be administered the AAV particles ofthe present disclosure at a dose of 1.5×10¹² vector genomes at a volumeof 900 ul per putamen.

In one embodiment, a subject may be administered the AAV particles ofthe present disclosure at a dose of 7.5×10¹¹ vector genomes at a volumeof 450 ul per putamen.

In one embodiment, a subject may be administered the AAV particles witha bilateral surgical infusion into at least one putamen using aposterior (i.e., back of the head) surgical delivery approach. Thenumber of posterior bilateral surgical infusions may be one or more suchas, but not limited to, 1 infusion, 2 infusions, 3 infusions, 4infusions or more than 4 infusions. As a non-limiting example, the AAVparticles are delivered in the left putamen with one posterior bilateralsurgical infusion. As a non-limiting example, the AAV particles aredelivered in the right putamen with one posterior bilateral surgicalinfusion. As a non-limiting example, the AAV particles are delivered inthe left putamen with two posterior bilateral surgical infusions. As anon-limiting example, the AAV particles are delivered in the rightputamen with two posterior bilateral surgical infusions. As anon-limiting example, the AAV particles are delivered in the right andleft putamen with two posterior bilateral surgical infusions. As anon-limiting example, the AAV particles are delivered in the leftputamen with three posterior bilateral surgical infusions. As anon-limiting example, the AAV particles are delivered in the rightputamen with three posterior bilateral surgical infusions. As anon-limiting example, the AAV particles are delivered in the right andleft putamen with three posterior bilateral surgical infusions. As anon-limiting example, the AAV particles are delivered in the leftputamen with four posterior bilateral surgical infusions. As anon-limiting example, the AAV particles are delivered in the rightputamen with four posterior bilateral surgical infusions. As anon-limiting example, the AAV particles are delivered in the right andleft putamen with four posterior bilateral surgical infusions.

In one embodiment, a subject may be administered the AAV particles witha bilateral surgical infusion into at least one putamen using atransfrontal (i.e., top of the head) surgical delivery approach. Thenumber of bilateral surgical infusions may be two or more such as, butnot limited to, 2 infusions, 3 infusions, 4 infusions or more than 4infusions. As a non-limiting example, the AAV particles are delivered inthe left putamen with 2 posterior bilateral surgical infusions. As anon-limiting example, the AAV particles are delivered in the rightputamen with 2 posterior bilateral surgical infusions. As a non-limitingexample, the AAV particles are delivered in the left and right putamenwith 2 posterior bilateral surgical infusions.

Delivery to the SNpc and VTA or STN

In one embodiment, a subject may be administered the AAV particles ofthe present disclosure safely delivered to substantia nigra parscompacta (SNpc) and ventral tegmental area (VTA) via bilateralinfusions, or alternatively, intrastriatally (into the caudate nucleusand putamen), or into the subthalamic nucleus (STN).

Delivery, Dose and Regimen

The present disclosure provides methods of administering AAV particlesin accordance with the disclosure to a subject in need thereof. Thepharmaceutical, diagnostic, or prophylactic AAV particles andcompositions of the present disclosure may be administered to a subjectusing any amount and any route of administration effective forpreventing, treating, managing, or diagnosing diseases, disorders and/orconditions. The exact amount required will vary from subject to subject,depending on the species, age, and general condition of the subject, theseverity of the disease, the particular composition, its mode ofadministration, its mode of activity, and the like. The subject may be ahuman, a mammal, or an animal. Compositions in accordance with thedisclosure are typically formulated in unit dosage form for ease ofadministration and uniformity of dosage. It will be understood, however,that the total daily usage of the compositions of the present disclosuremay be decided by the attending physician within the scope of soundmedical judgment. The specific therapeutically effective,prophylactically effective, or appropriate diagnostic dose level for anyparticular individual will depend upon a variety of factors includingthe disorder being treated and the severity of the disorder; theactivity of the specific payload employed; the specific compositionemployed; the age, body weight, general health, sex and diet of thepatient; the time of administration, route of administration, and rateof excretion of the specific AAV particle employed; the duration of thetreatment; drugs used in combination or coincidental with the specificAAV particle employed; and like factors well known in the medical arts.

In one embodiment, delivery of the AAV particles of the presentdisclosure results in minimal serious adverse events (SAEs) as a resultof the delivery of the AAV particles.

In one embodiment, the AAV particle may be delivered a multi-doseregimen. The multi-dose regimen may be 2, 3, 4, 5, 6, 7, 8, 9, 10 ormore than 10 doses.

In one embodiment, the AAV particle may be delivered to a subject via amulti-site route of administration. A subject may be administered theAAV particle at 2, 3, 4, 5 or more than 5 sites.

Dosage Levels

In certain embodiments, AAV particle pharmaceutical compositions inaccordance with the present disclosure may be administered at dosagelevels sufficient to deliver from about 0.0001 mg/kg to about 100 mg/kg,from about 0.001 mg/kg to about 0.05 mg/kg, from about 0.005 mg/kg toabout 0.05 mg/kg, from about 0.001 mg/kg to about 0.005 mg/kg, fromabout 0.05 mg/kg to about 0.5 mg/kg, from about 0.01 mg/kg to about 50mg/kg, from about 0.1 mg/kg to about 40 mg/kg, from about 0.5 mg/kg toabout 30 mg/kg, from about 0.01 mg/kg to about 10 mg/kg, from about 0.1mg/kg to about 10 mg/kg, or from about 1 mg/kg to about 25 mg/kg, ofsubject body weight per day, one or more times a day, to obtain thedesired therapeutic, diagnostic, or prophylactic, effect. It will beunderstood that the above dosing concentrations may be converted to vgor viral genomes per kg or into total viral genomes administered by oneof skill in the art.

In certain embodiments, AAV particle pharmaceutical compositions inaccordance with the present disclosure may be administered at about 10to about 600 μl/site, 50 to about 500 μl/site, 100 to about 400 μl/site,120 to about 300 μl/site, 140 to about 200 μl/site, about 160 μl/site.As non-limiting examples, AAV particles may be administered at 50μl/site and/or 150 μl/site.

In one embodiment, delivery of the compositions in accordance with thepresent disclosure to cells comprises a rate of delivery defined by[VG/hour=mL/hour*VG/mL] wherein VG is viral genomes, VG/mL iscomposition concentration, and mL/hour is rate of prolonged delivery.

In one embodiment, delivery of compositions in accordance with thepresent disclosure to cells may comprise a total concentration persubject between about 1×10⁶ VG and about 1×10¹⁶ VG. In some embodiments,delivery may comprise a composition concentration of about 1×10⁶, 2×10⁶,3×10⁶, 4×10⁶, 5×10⁶, 6×10⁶, 7×10⁶, 8×10⁶, 9×10⁶, 1×10⁷, 2×10⁷, 3×10⁷,4×10⁷, 5×10⁷, 6×10⁷, 7×10⁷, 8×10⁷, 9×10⁷, 1×10⁸, 2×10⁸, 3×10⁸, 4×10⁸,5×10⁸, 6×10⁸, 7×10⁸, 8×10⁸, 9×10⁸, 1×10⁹, 2×10⁹, 3×10⁹, 4×10⁹, 5×10⁹,6×10⁹, 7×10⁹, 8×10⁹, 9×10⁹, 1×10¹⁰, 2×10¹⁰, 3×10¹⁰, 4×10¹⁰, 5×10¹⁰,6×10¹⁰, 7×10¹⁰, 8×10¹⁰, 9×10¹⁰, 1×10¹¹, 2×10¹¹, 2.1×10¹¹, 2.2×10¹¹,2.3×10¹¹, 2.4×10¹¹, 2.5×10¹¹, 2.6×10¹¹, 2.7×10¹¹, 2.8×10¹¹, 2.9×10¹¹,3×10¹¹, 4×10¹¹, 5×10¹¹, 6×10¹¹, 7×10¹¹, 7.1×10¹¹, 7.2×10¹¹, 7.3×10¹¹,7.4×10¹¹, 7.5×10¹¹, 7.6×10¹¹, 7.7×10¹¹, 7.8×10¹¹, 7.9×10¹¹, 8×10¹¹,9×10¹¹, 1×10¹², 1.1×10¹², 1.2×10¹², 1.3×10¹², 1.4×10¹², 1.5×10¹²,1.6×10¹², 1.7×10¹², 1.8×10¹², 1.9×10¹², 2×10¹², 3×10¹², 4×10¹²,4.1×10¹², 4.2×10¹², 4.3×10¹², 4.4×10¹², 4.5×10¹², 4.6×10¹², 4.7×10¹²,4.8×10¹², 4.9×10¹², 5×10¹², 6×10¹², 7×10¹², 8×10¹², 8.1×10¹², 8.2×10¹²,8.3×10¹², 8.4×10¹², 8.5×10¹², 8.6×10¹², 8.7×10¹², 8.8×10¹², 8.9×10¹²,9×10¹², 1×10¹³, 2×10¹³, 3×10¹³, 4×10¹³, 5×10¹³, 6×10¹³, 6.7×10¹³,7×10¹³, 8×10¹³, 9×10¹³, 1×10¹⁴, 2×10¹⁴, 3×10¹⁴, 4×10¹⁴, 5×10¹⁴, 6×10¹⁴,7×10¹⁴, 8×10¹⁴, 9×10¹⁴, 1×10¹⁵, 2×10¹⁵, 3×10¹⁵, 4×10¹⁵, 5×10¹⁵, 6×10¹⁵,7×10¹⁵, 8×10¹⁵, 9×10¹⁵, or 1×10¹⁶VG/subject. In one embodiment, theconcentration of the AAV particle in the composition is 1×10¹³VG/subject. In one embodiment, the concentration of the AAV particle inthe composition is 3×10¹² VG/subject. As a non-limiting example, thecomposition administered to the subject has a concentration of about3×10¹¹ VG/subject. As a non-limiting example, the compositionadministered to the subject has a concentration of about 9×10¹¹VG/subject. In one embodiment, the concentration of the AAV particle inthe composition is 2.3×10¹¹ VG/subject. In one embodiment, theconcentration of the AAV particle in the composition is 7.2×10¹¹VG/subject. In one embodiment, the concentration of the AAV particle inthe composition is 7.5×10¹¹ VG/subject. In one embodiment, theconcentration of the AAV particle in the composition is 1.4×10¹²VG/subject. In one embodiment, the concentration of the AAV particle inthe composition is 4.8×10¹² VG/subject. In one embodiment, theconcentration of the AAV particle in the composition is 8.8×10¹²VG/subject. In one embodiment, the concentration of the AAV particle inthe composition is 2.3×10¹² VG/subject.

In one embodiment, delivery of compositions in accordance with thepresent disclosure to cells may comprise a total concentration persubject between about 1×10⁶VG/kg and about 1×10¹⁶ VG/kg. In someembodiments, delivery may comprise a composition concentration of about1×10⁶, 2×10⁶, 3×10⁶, 4×10⁶, 5×10⁶, 6×10⁶, 7×10⁶, 8×10⁶, 9×10⁶, 1×10⁷,2×10⁷, 3×10⁷, 4×10⁷, 5×10⁷, 6×10⁷, 7×10⁷, 8×10⁷, 9×10⁷, 1×10⁸, 2×10⁸,3×10⁸, 4×10⁸, 5×10⁸, 6×10⁸, 7×10⁸, 8×10⁸, 9×10⁸, 1×10⁹, 2×10⁹, 3×10⁹,4×10⁹, 5×10⁹, 6×10⁹, 7×10⁹, 8×10⁹, 9×10⁹, 1×10¹⁰, 2×10¹⁰, 3×10¹⁰,4×10¹⁰, 5×10¹⁰, 6×10¹⁰, 7×10¹⁰, 8×10¹⁰, 9×10¹⁰, 1×10¹¹, 2×10¹¹,2.1×10¹¹, 2.2×10¹¹, 2.3×10¹¹, 2.4×10¹¹, 2.5×10¹¹, 2.6×10¹¹, 2.7×10¹¹,2.8×10¹¹, 2.9×10¹¹, 3×10¹¹, 4×10¹¹, 5×10¹¹, 6×10¹¹, 7×10¹¹, 7.1×10¹¹,7.2×10¹¹, 7.3×10¹¹, 7.4×10¹¹, 7.5×10¹¹, 7.6×10¹¹, 7.7×10¹¹, 7.8×10¹¹,7.9×10¹¹, 8×10¹¹, 9×10¹¹, 1×10¹², 1.1×10¹², 1.2×10¹², 1.3×10¹²,1.4×10¹², 1.5×10¹², 1.6×10¹², 1.7×10¹², 1.8λ10¹², 1.9λ10¹², 2×10¹²,3×10¹², 4×10¹², 4.1×10¹², 4.2×10¹², 4.3×10¹², 4.4×10¹², 4.5×10¹²,4.6×10¹², 4.7×10¹², 4.8×10¹², 4.9×10¹², 5×10¹², 6×10¹², 7×10¹², 8×10¹²,8.1×10¹², 8.2×10¹², 8.3×10¹², 8.4×10¹², 8.5×10¹², 8.6×10¹², 8.7×10¹²,8.8×10¹², 8.9×10¹², 9×10¹², 1×10¹³, 2×10¹³, 3×10¹³, 4×10¹³, 5×10¹³,6×10¹³, 6.7×10¹³, 7×10¹³, 8×10¹³, 9×10¹³, 1×10¹⁴, 2×10¹⁴, 3×10¹⁴,4×10¹⁴, 5×10¹⁴, 6×10¹⁴, 7×10¹⁴, 8×10¹⁴, 9×10¹⁴, 1×10¹⁵, 2×10¹⁵, 3×10¹⁵,4×10¹⁵, 5×10¹⁵, 6×10¹⁵, 7×10¹⁵, 8×10¹⁵, 9×10¹⁵, or 1×10¹⁶ VG/kg.

In one embodiment, delivery of AAV particles to cells of the centralnervous system (e.g., parenchyma) may comprise a total dose betweenabout 1×10⁶VG and about 1×10¹⁶VG. In some embodiments, delivery maycomprise a total dose of about 1×10⁶, 2×10⁶, 3×10⁶, 4×10⁶, 5×10⁶, 6×10⁶,7×10⁶, 8×10⁶, 9×10⁶, 1×10⁷, 2×10⁷, 3×10⁷, 4×10⁷, 5×10⁷, 6×10⁷, 7×10⁷,8×10⁷, 9×10⁷, 1×10⁸, 2×10⁸, 3×10⁸, 4×10⁸, 5×10⁸, 6×10⁸, 7×10⁸, 8×10⁸,9×10⁸, 1×10⁹, 2×10⁹, 3×10⁹, 4×10⁹, 5×10⁹, 6×10⁹, 7×10⁹, 8×10⁹, 9×10⁹,1×10¹⁰, 1.9×10¹⁰, 2×10¹⁰, 3×10¹⁰, 3.73×10¹⁰, 4×10¹⁰, 5×10¹⁰, 6×10¹⁰,7×10¹⁰, 8×10¹⁰, 9×10¹⁰, 1×10¹¹, 2×10¹¹, 2.5×10¹¹, 3×10¹¹, 4×10¹¹,5×10¹¹, 6×10¹¹, 7×10¹¹, 8×10¹¹, 9×10¹¹, l×10¹², 2×10¹², 3×10¹², 4×10¹²,5×10¹², 6×10¹², 7×10¹², 8×10¹², 9×10¹², l×10¹³, 2×10¹³, 3×10¹³, 4×10¹³,5×10¹³, 6×10¹³, 7×10¹³, 8×10¹³, 9×10¹³, l×10¹⁴, 2×10¹⁴, 3×10¹⁴, 4×10¹⁴,5×10¹⁴, 6×10¹⁴, 7×10¹⁴, 8×10¹⁴, 9×10¹⁴, 1×10¹⁵, 2×10¹⁵, 3×10¹⁵, 4×10¹⁵,5×10¹⁵, 6×10¹⁵, 7×10¹⁵, 8×10¹⁵, 9×10¹⁵, or 1×10¹⁶ VG. As a non-limitingexample, the total dose is l×10¹¹ VG. As another non-limiting example,the total dose is 2.1×10¹² VG.

In one embodiment, about 10⁵ to 10⁶ viral genome (unit) may beadministered per dose.

In one embodiment, delivery of the compositions in accordance with thepresent disclosure to cells may comprise a total concentration betweenabout 1×10⁶VG/mL and about 1×10¹⁶VG/mL. In some embodiments, deliverymay comprise a composition concentration of about 1×10⁶, 2×10⁶, 3×10⁶,4×10⁶, 5×10⁶, 6×10⁶, 7×10⁶, 8×10⁶, 9×10⁶, 1×10⁷, 2×10⁷, 3×10⁷, 4×10⁷,5×10⁷, 6×10⁷, 7×10⁷, 8×10⁷, 9×10⁷, 1×10⁸, 2×10⁸, 3×10⁸, 4×10⁸, 5×10⁸,6×10⁸, 7×10⁸, 8×10⁸, 9×10⁸, 1×10⁹, 2×10⁹, 3×10⁹, 4×10⁹, 5×10⁹, 6×10⁹,7×10⁹, 8×10⁹, 9×10⁹, 1×10¹⁰, 2×10¹⁰, 3×10¹⁰, 4×10¹⁰, 5×10¹⁰, 6×10¹⁰,7×10¹⁰, 8×10¹⁰, 9×10¹⁰, 1×10¹¹, 2×10¹¹, 3×10¹¹, 4×10¹¹, 5×10¹¹, 6×10¹¹,7×10¹¹, 8×10¹¹, 9×10¹¹, 1×10¹², 1.1×10¹², 1.2×10¹², 1.3×10¹², 1.4×10¹²,1.5×10¹², 1.6×10¹², 1.7×10¹², 1.8×10¹², 1.9×10¹², 2×10¹², 2.1×10¹²,2.2×10¹², 2.3×10¹², 2.4×10¹², 2.5×10¹², 2.6×10¹², 2.7×10¹², 2.8×10¹²,2.9×10¹², 3×10¹², 3.1×10¹², 3.2×10¹², 3.3×10¹², 3.4×10¹², 3.5×10¹²,3.6×10¹², 3.7×10¹², 3.8×10¹², 3.9×10¹², 4×10¹², 4.1×10¹², 4.2×10¹²,4.3×10¹², 4.4×10¹², 4.5×10¹², 4.6×10¹², 4.7×10¹², 4.8×10¹², 4.9×10¹²,5×10¹², 6×10¹², 7×10¹², 8×10¹², 9×10¹², 1×10¹³, 2×10¹³, 3×10¹³, 4×10¹³,5×10¹³, 6×10¹³, 6.7×10¹³, 7×10¹³, 8×10¹³, 9×10¹³, 1×10¹⁴ 2×10¹⁴ 3×10¹⁴4×10¹⁴ 5×10¹⁴ 6×10¹⁴ 7×10¹⁴ 8×10¹⁴ 9×10¹⁴ 1×10¹⁵ 2×10¹⁵ 3×10¹⁵ 4×10¹⁵5×10¹⁵, 6×10¹⁵, 7×10¹⁵, 8×10¹⁵, 9×10¹⁵, or 1×10¹⁶VG/mL. In oneembodiment, the concentration of the AAV particle in the composition is1×10¹³VG/mL. In one embodiment, the concentration of the AAV particle inthe composition is 3×10¹²VG/mL. In one embodiment, the concentration ofthe AAV particle in the composition is 1.1×10¹²VG/mL. In one embodiment,the concentration of the AAV particle in the composition is3.7×10¹²VG/mL. In one embodiment, the concentration of the AAV particlein the composition is 8×10¹¹ VG/mL. In one embodiment, the concentrationof the AAV particle in the composition is 2.6×10¹²VG/mL. In oneembodiment, the concentration of the AAV particle in the composition is4.9×10¹²VG/mL. In one embodiment, the concentration of the AAV particlein the composition is 0.8×10¹²VG/mL. In one embodiment, theconcentration of the AAV particle in the composition is 0.83×10¹²VG/mL.In one embodiment, the concentration of the AAV particle in thecomposition is the maximum final dose which can be contained in a vial.

In one embodiment, delivery of AAV particles to cells of the centralnervous system (e.g., parenchyma) may comprise a compositionconcentration between about 1×10⁶VG/mL and about 1×10¹⁶VG/mL. In someembodiments, delivery may comprise a composition concentration of about1×10⁶ 2×10⁶ 3×10⁶ 4×10⁶ 5×10⁶ 6×10⁶ 7×10⁶ 8×10⁶ 9×10⁶ 1×10⁷ 2×10⁷ 3×10⁷4×10⁷ 5×10⁷ 6×10⁷, 7×10⁷, 8×10⁷, 9×10⁷, 1×10⁸, 2×10⁸, 3×10⁸, 4×10⁸,5×10⁸, 6×10⁸, 7×10⁸, 8×10⁸, 9×10⁸, 1×10⁹, 2×10⁹, 3×10⁹, 4×10⁹, 5×10⁹,6×10⁹, 7×10⁹, 8×10⁹, 9×10⁹, 1×10¹⁰, 2×10¹⁰, 3×10¹⁰, 4×10¹⁰, 5×10¹⁰,6×10¹⁰, 7×10¹⁰, 8×10¹⁰, 9×10¹⁰, 1×10¹¹, 2×10¹¹, 3×10¹¹, 4×10¹¹, 5×10¹¹,6×10¹¹, 7×10¹¹, 8×10¹¹, 9×10¹¹, 1×10¹², 2×10¹², 3×10¹², 4×10¹², 5×10¹²,6×10¹², 7×10¹², 8×10¹², 9×10¹², 1×10¹³, 2×10¹³, 3×10¹³, 4×10¹³, 5×10¹³,6×10¹³, 7×10¹³, 8×10¹³, 9×10¹³, 1×10¹⁴, 2×10¹⁴, 3×10¹⁴, 4×10¹⁴, 5×10¹⁴,6×10¹⁴, 7×10¹⁴, 8×10¹⁴, 9×10¹⁴, 1×10¹⁵, 2×10¹⁵, 3×10¹⁵, 4×10¹⁵, 5×10¹⁵,6×10¹⁵ 7×10¹⁵ 8×10¹⁵, 9×10¹⁵, or 1×10¹⁶VG/mL. In one embodiment, thedelivery comprises a composition concentration of 1×10¹³VG/mL. In oneembodiment, the delivery comprises a composition concentration of2.1×10¹²VG/mL.

Regimen

The desired dosage of the AAV particles of the present disclosure may bedelivered only once, three times a day, two times a day, once a day,every other day, every third day, every week, every two weeks, everythree weeks, or every four weeks. In certain embodiments, the desireddosage may be delivered using multiple administrations (e.g., two,three, four, five, six, seven, eight, nine, ten, eleven, twelve,thirteen, fourteen, or more administrations). When multipleadministrations are employed, split dosing regimens such as thosedescribed herein may be used. As used herein, a “split dose” is thedivision of “single unit dose” or total daily dose into two or moredoses, e.g., two or more administrations of the “single unit dose”. Asused herein, a “single unit dose” is a dose of any therapeuticadministered in one dose/at one time/single route/single point ofcontact, i.e., single administration event.

The desired dosage of the AAV particles of the present disclosure may beadministered as a “pulse dose” or as a “continuous flow”. As usedherein, a “pulse dose” is a series of single unit doses of anytherapeutic administered with a set frequency over a period of time. Asused herein, a “continuous flow” is a dose of therapeutic administeredcontinuously for a period of time in a single route/single point ofcontact, i.e., continuous administration event. A total daily dose, anamount given or prescribed in 24 hour period, may be administered by anyof these methods, or as a combination of these methods, or by any othermethods suitable for a pharmaceutical administration.

In one embodiment, delivery of the AAV particles of the presentdisclosure to a subject provides regulating activity of AADC in asubject. The regulating activity may be an increase in the production ofAADC in a subject. The regulating activity can be for at least 1 month,2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9months, 10 months, 11 months, 1 year, 13 months, 14 months, 15 months,16 months, 17 months, 18 months, 19 months, 20 months, 20 months, 21months, 22 months, 23 months, 2 years, 3 years, 4 years, 5 years, 6years, 7 years, 8 years, 9 years, 10 years or more than 10 years.

In some embodiments, the AAV particle of the present disclosure may beadministered to a subject using a single dose, one-time treatment. Thedose of the one-time treatment may be administered by any methods knownin the art and/or described herein. As used herein, a “one-timetreatment” refers to a composition which is only administered one time.If needed, a booster dose may be administered to the subject to ensurethe appropriate efficacy is reached. A booster may be administered 1month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8months, 9 months, 10 months, 11 months, 12 months, 1 year, 13 months, 14months, 15 months, 16 months, 17 months, 18 months, 19 months, 20months, 21 months, 22 months, 23 months, 24 months, 2 years, 3 years, 4years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, or morethan 10 years after the one-time treatment.

Delivery Methods

In one embodiment, the AAV particles or pharmaceutical compositions ofthe present disclosure may be administered or delivered using themethods for treatment of disease described in U.S. Pat. No. 8,999,948,or International Publication No. WO2014178863, the contents of which areherein incorporated by reference in their entirety.

In one embodiment, the AAV particles or pharmaceutical compositions ofthe present disclosure may be administered or delivered using themethods for delivering gene therapy in Alzheimer's Disease or otherneurodegenerative conditions as described in US Application No.20150126590, the contents of which are herein incorporated by referencein their entirety.

In one embodiment, the AAV particles or pharmaceutical compositions ofthe present disclosure may be administered or delivered using themethods for delivery of a CNS gene therapy as described in U.S. Pat.Nos. 6,436,708, and 8,946,152, and International Publication No.WO2015168666, the contents of which are herein incorporated by referencein their entirety.

In one embodiment, the AAV particle comprising an AADC polynucleotidemay be administered or delivered using the methods for the delivery ofAAV virions described in European Patent Application No. EP1857552, thecontents of which are herein incorporated by reference in its entirety.

In one embodiment, the AAV particle or pharmaceutical compositions ofthe present disclosure may be administered or delivered using themethods for delivering proteins using AAV particles described inEuropean Patent Application No. EP2678433, the contents of which areherein incorporated by reference in their entirety.

In one embodiment, the AAV particle comprising an AADC polynucleotidemay be administered or delivered using the methods for delivering DNAmolecules using AAV particles described in U.S. Pat. No. 5,858,351, thecontents of which are herein incorporated by reference in its entirety.

In one embodiment, the AAV particle or pharmaceutical compositions ofthe present disclosure may be administered or delivered using themethods for delivering DNA to the bloodstream described in U.S. Pat. No.6,211,163, the contents of which are herein incorporated by reference intheir entirety.

In one embodiment, the AAV particle or pharmaceutical compositions ofthe present disclosure may be administered or delivered using themethods for delivering a payload to the central nervous system describedin U.S. Pat. No. 7,588,757, the contents of which are hereinincorporated by reference in their entirety.

In one embodiment, the AAV particle may be administered or deliveredusing the methods for delivering AAV virions described in U.S. Pat. No.6,325,998, the contents of which are herein incorporated by reference inits entirety.

In one embodiment, the AAV particle or pharmaceutical compositions ofthe present disclosure may be administered or delivered using themethods for delivering a payload described in U.S. Pat. No. 8,283,151,the contents of which are herein incorporated by reference in theirentirety.

In one embodiment, the AAV particle or pharmaceutical compositions ofthe present disclosure may be administered or delivered using themethods for delivering a payload using a glutamic acid decarboxylase(GAD) delivery vector described in International Patent Publication No.WO2001089583, the contents of which are herein incorporated by referencein their entirety.

In one embodiment, the AAV particle or pharmaceutical compositions ofthe present disclosure may be administered or delivered using themethods for delivering a payload to neural cells described inInternational Patent Publication No. WO2012057363, the contents of whichare herein incorporated by reference in their entirety.

Delivery to Cells

The present disclosure provides a method of delivering to a cell ortissue any of the above-described AAV particles, comprising contactingthe cell or tissue with said AAV particle or contacting the cell ortissue with a formulation comprising said AAV particle, or contactingthe cell or tissue with any of the described compositions, includingpharmaceutical compositions. The method of delivering the AAV particleto a cell or tissue can be accomplished in vitro, ex vivo, or in vivo.

Delivery to Subjects

The present disclosure additionally provides a method of delivering to asubject, including a mammalian subject, any of the above-described AAVparticles comprising administering to the subject said AAV particle, oradministering to the subject a formulation comprising said AAV particle,or administering to the subject any of the described compositions,including pharmaceutical compositions.

Combinations

The AAV particles may be used in combination with one or more othertherapeutic, prophylactic, research or diagnostic agents. By “incombination with,” it is not intended to imply that the agents must beadministered at the same time and/or formulated for delivery together,although these methods of delivery are within the scope of the presentdisclosure. Compositions can be administered concurrently with, priorto, or subsequent to, one or more other desired therapeutics or medicalprocedures. In general, each agent will be administered at a dose and/oron a time schedule determined for that agent. In some embodiments, thepresent disclosure encompasses the delivery of pharmaceutical,prophylactic, research, or diagnostic compositions in combination withagents that may improve their bioavailability, reduce and/or modifytheir metabolism, inhibit their excretion, and/or modify theirdistribution within the body.

In one embodiment, the AAV particles described herein may beadministered to a subject who is also undergoing levodopa therapy. As anon-limiting example, the subject may have a positive response tolevodopa therapy and at least one symptom of PD is reduced. As anothernon-limiting example, the subject may have a response to levodopatherapy where the symptoms of PD experienced by the subject are stable.As yet another non-limiting example, the subject may have a negativeresponse to levodopa therapy where the symptoms of PD experienced by thesubject are increasing.

In one embodiment, the dose of levodopa administered to the subjectprior to the AAV articles is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more than 25 mg/kg. Asa non-limiting example, the dose is 3 mg/kg. As another non-limitingexample, the dose is 10 mg/kg. As yet another non-limiting example, thedose is 20 mg/kg. The subject's response (e.g., behavioral response) tolevodopa may be assessed prior to administration of the AAV particles.Additionally, the subject may be administered levodopa again after theadministration of the AADC polynucleotides (e.g., 1 week, 2, weeks, 3weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7months, 8 months, 9 months, 10 months, 11 months, 1 year or more than 1year after the administration of AAV particles). The behavioral responsecan be re-assessed and compared to the initial response to determine theeffects of the AAV particles. The subject may have 1%, 2%, 3%, 4%, 5%,6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100% behavioral improvement.

In one embodiment, Levodopa may be administered multiple times after theadministration of the AAV particles. Levodopa may be administered on arepeating schedule (e.g., every 5 days, weekly, every 10 days, every 15days, every 30 days, monthly, bimonthly, every 3 months, every 4 months,every 5 months, every 6 months, every 7 months, every 8 months, every 9months, every 10 months, every 11 months or yearly) or as symptomsarise. As a non-limiting example, 3 years post administration of AADCpolynucleotides a subject may have 1-10%, 5-15%, 10-20%, 15-30%, 20-40%,25-50%, 30-50%, 40-50%, 40-60%, 50-70%, 50-80%, 60-70%, 60-75%, 60-80%,60-90%, 70-80%, 70-90%, 75-90%, 80-90%, 90-100% of the striatal neuronswithin the infused region of the putamen to be AADC-immunoreactive. As anon-limiting example, 6 years post administration of AAV particles asubject may have 1-10%, 5-15%, 10-20%, 15-30%, 20-40%, 25-50%, 30-50%,40-50%, 40-60%, 50-70%, 50-80%, 60-70%, 60-75%, 60-80%, 60-90%, 70-80%,70-90%, 75-90%, 80-90%, 90-100% of the striatal neurons within theinfused region of the putamen to be AADC-immunoreactive. As anon-limiting example, 9 years post administration of AADCpolynucleotides a subject may have 1-10%, 5-15%, 10-20%, 15-30%, 20-40%,25-50%, 30-50%, 40-50%, 40-60%, 50-70%, 50-80%, 60-70%, 60-75%, 60-80%,60-90%, 70-80%, 70-90%, 75-90%, 80-90%, 90-100% of the striatal neuronswithin the infused region of the putamen to be AADC-immunoreactive.

In one embodiment, a subject who may be administered the AAV particlesdescribed herein have a documented response to levodopa therapy but havemedically refractory fluctuations and are considered good surgicalcandidates. The determination if a subject is a good surgical candidatemay be made by the physician treating the subject for PD or thephysician administering the AAV particles who takes into considerationthe overall risk to benefit ratio for the patient for the surgicalintervention required for delivery of the AAV particles.

Measurement of Expression

Expression of payloads from viral genomes may be determined usingvarious methods known in the art such as, but not limited toimmunochemistry (e.g., IHC), in situ hybridization (ISH), enzyme-linkedimmunosorbent assay (ELISA), affinity ELISA, ELISPOT, flow cytometry,immunocytology, surface plasmon resonance analysis, kinetic exclusionassay, liquid chromatography-mass spectrometry (LCMS), high-performanceliquid chromatography (HPLC), BCA assay, immunoelectrophoresis, Westernblot, SDS-PAGE, protein immunoprecipitation, and/or PCR.

The pharmaceutical compositions of AAV particles described herein may becharacterized by one or more of bioavailability, therapeutic windowand/or volume of distribution.

Bioavailability

The AAV particles, when formulated into a composition with a deliveryagent as described herein, can exhibit an increase in bioavailability ascompared to a composition lacking a delivery agent as described herein.As used herein, the term “bioavailability” refers to the systemicavailability of a given amount of AAV particle or expressed payloadadministered to a mammal. Bioavailability can be assessed by measuringthe area under the curve (AUC) or the maximum serum or plasmaconcentration (C_(max)) of the composition following. AUC is adetermination of the area under the curve plotting the serum or plasmaconcentration of a compound (e.g., AAV particles or expressed payloads)along the ordinate (Y-axis) against time along the abscissa (X-axis).Generally, the AUC for a particular compound can be calculated usingmethods known to those of ordinary skill in the art and as described inG. S. Banker, Modern Pharmaceutics, Drugs and the PharmaceuticalSciences, v. 72, Marcel Dekker, New York, Inc., 1996, the contents ofwhich are herein incorporated by reference in its entirety.

The C_(max) value is the maximum concentration of the AAV particle orexpressed payload achieved in the serum or plasma of a mammal followingadministration of the AAV particle to the mammal. The C_(max) value ofcan be measured using methods known to those of ordinary skill in theart. The phrases “increasing bioavailability” or “improving thepharmacokinetics,” as used herein mean that the systemic availability ofa first AAV particle or expressed payload, measured as AUC, C_(max), orC_(min) a mammal is greater, when co-administered with a delivery agentas described herein, than when such co-administration does not takeplace. In some embodiments, the bioavailability can increase by at leastabout 2%, at least about 5%, at least about 10%, at least about 15%, atleast about 20%, at least about 25%, at least about 30%, at least about35%, at least about 40%, at least about 45%, at least about 50%, atleast about 55%, at least about 60%, at least about 65%, at least about70%, at least about 75%, at least about 80%, at least about 85%, atleast about 90%, at least about 95%, or about 100%.

Therapeutic Window

As used herein “therapeutic window” refers to the range of plasmaconcentrations, or the range of levels of therapeutically activesubstance at the site of action, with a high probability of eliciting atherapeutic effect. In some embodiments, the therapeutic window of theAAV particle as described herein can increase by at least about 2%, atleast about 5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, atleast about 40%, at least about 45%, at least about 50%, at least about55%, at least about 60%, at least about 65%, at least about 70%, atleast about 75%, at least about 80%, at least about 85%, at least about90%, at least about 95%, or about 100%.

Volume of Distribution

As used herein, the term “volume of distribution” refers to the fluidvolume that would be required to contain the total amount of the drug inthe body at the same concentration as in the blood or plasma: V_(dist)equals the amount of drug in the body/concentration of drug in blood orplasma. For example, for a 10 mg dose and a plasma concentration of 10mg/L, the volume of distribution would be 1 liter. The volume ofdistribution reflects the extent to which the drug is present in theextravascular tissue. A large volume of distribution reflects thetendency of a compound to bind to the tissue components compared withplasma protein binding. In a clinical setting, V_(dist) can be used todetermine a loading dose to achieve a steady state concentration. Insome embodiments, the volume of distribution of the AAV particles asdescribed herein can decrease at least about 2%, at least about 5%, atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, at least about 50%, at least about 55%, at least about60%, at least about 65%, at least about 70%.

Biological Effect

In one embodiment, the biological effect of the AAV particles deliveredto the animals may be categorized by analyzing the payload expression inthe animals. The payload expression may be determined from analyzing abiological sample collected from a mammal administered the AAV particlesof the present disclosure. For example, a protein expression of 50-200pg/ml for the protein encoded by the AAV particles delivered to themammal may be seen as a therapeutically effective amount of protein inthe mammal.

IV. Methods and Uses of the Compositions CNS Diseases

The polynucleotides of the present disclosure may be used in thetreatment, prophylaxis or amelioration of any disease or disordercharacterized by aberrant or undesired target expression. In oneembodiment, the disclosure relates to AAV particles for use in thetreatment of Parkinson's disease.

In some embodiments, the AAV particles may be used in the treatment,prophylaxis or amelioration of any disease or disorder characterized byaberrant or undesired target expression wherein the payload, i.e. AADC,is swapped for an alternate payload.

The present disclosure provides a method for treating a disease,disorder and/or condition in a mammalian subject, including a humansubject, comprising administering to the subject AAV particles describedherein.

In one embodiment, the disease, disorder and/or condition is aneurological disease, disorder and/or condition. The CNS diseases may bediseases that affect any component of the brain (including the cerebralhemispheres, diencephalon, brain stem, and cerebellum) or the spinalcord.

In some embodiments, viral particles of the present disclosure, throughdelivery of a functional payload that is a therapeutic product that canmodulate the level or function of a gene product in the CNS, may be usedto treat a neurodegenerative diseases and/or diseases or disorders thatare characteristic with neurodegeneration, neuromuscular diseases,lysosomal diseases, trauma, bone marrow injuries, pain (includingneuropathic pain), cancers of the nervous system, demyelinatingdiseases, autoimmune diseases of the nervous system, neurotoxicsyndromes, sleeping disorders, genetic brain disorders and developmentalCNS disorders. A functional payload may alleviate or reduce symptomsthat result from abnormal level and/or function of a gene product (e.g.,an absence or defect in a protein) in a subject in need thereof or thatotherwise confers a benefit to a CNS disorder in a subject in needthereof.

As non-limiting examples, therapeutic products delivered by AAVparticles of the present disclosure may include, but are not limited to,growth and trophic factors, cytokines, hormones, neurotransmitters,enzymes, anti-apoptotic factors, angiogenic factors, and any proteinknown to be mutated in pathological disorders such as the “survival ofmotor neuron” protein (SMN); antisense RNA or RNAi targeting messengerRNAs coding for proteins having a therapeutic interest in any of CNSdiseases discussed herein; or microRNAs that function in gene silencingand post-transcriptionally regulation of gene expression in the CNS(e.g., brain specific Mir-128a, See Adlakha and Saini, Molecular cancer,2014, 13:33). For example, an RNAi targeting the superoxide dismutaseenzyme may be packaged by viral particles of the present disclosure, forthe treatment of ALS.

The growth and trophic factors may include, but are not limited tobrain-derived growth factor (BDNF), epidermal growth factor (EGF), basicFibroblast growth factor (bFGF), Ciliary neurotrophic factor (CNTF),corticotropin-releasing factor (CRF), Glial cell line derived growthfactor (GDNF), Insulin-like growth factor-1 (IGF-1), nerve growth factor(NGF), neurotrophin-3 (NT-3), neurotrophin-4 (NT-4), and vascularendothelial growth factor (VEGF). Cytokines may include interleukin-10(IL-10), interleukin-6, Interleukin-8, chemokine CXCL12 (SDF-1),TGF-beta, and Growth and differentiation factor (GDF-1/10).

In some embodiments, the neurological disorders may be neurodegenerativedisorders including, but not limited to, Alzheimer's Diseases (AD);Amyotrophic lateral sclerosis (ALS); Creutzfeldt-Jakob Disease (CJD);Huntingtin's disease (HD); Friedreich's ataxia (FA); Parkinson Disease(PD); Multiple System Atrophy (MSA); Spinal Muscular Atrophy (SMA),Multiple Sclerosis (MS); Primary progressive aphasia; Progressivesupranuclear palsy (PSP); Dementia; Brain Cancer, Degenerative NerveDiseases, Encephalitis, Epilepsy, Genetic Brain Disorders that causeneurodegeneration, Retinitis pigmentosa (RP), Head and BrainMalformations, Hydrocephalus, Stroke, Prion disease, Infantile neuronalceroid lipofuscinosis (INCL) (a neurodegenerative disease of childrencaused by a deficiency in the lysosomal enzyme palmitoyl proteinthioesterase-1 (PPT1)), and others.

In some embodiments, viral particles of the present disclosure may beused to treat diseases that are associated with impairments of thegrowth and development of the CNS, i.e., neurodevelopmental disorders.In some aspects, such neurodevelopmental disorders may be caused bygenetic mutations, including but not limited to, Fragile X syndrome(caused by mutations in FMR1 gene), Down syndrome (caused by trisomy ofchromosome 21), Rett syndrome, Williams syndrome, Angelman syndrome,Smith-Magenis syndrome, ATR-X syndrome, Barth syndrome, Immunedysfunction and/or infectious diseases during infancy such as Sydenham'schorea, Schizophrenia Congenital toxoplasmosis, Congenital rubellasyndrome, Metabolic disorders such as diabetes mellitus andphenylketonuria; nutritional defects and/or brain trauma, Autism andautism spectrum.

In some embodiments, viral particles of the present disclosure, may beused to treat a tumor in the CNS, including but not limited to, acousticneuroma, Astrocytoma (Grades I, II, III and IV), Chordoma, CNS Lymphoma,Craniopharyngioma, Gliomas (e.g., brain stem glioma, ependymoma, opticalnerve glioma, subependymoma), Medulloblastoma, Meningioma, Metastaticbrain tumors, Oligodendroglioma, Pituitary Tumors, Primitiveneuroectodermal (PNET), and Schwannoma.

In some embodiments, the neurological disorders may be functionalneurological disorders with motor and/or sensory symptoms which haveneurological origin in the CNS. As non-limiting examples, functionalneurological disorders may be chronic pain, seizures, speech problems,involuntary movements, and sleep disturbances.

In some embodiments, the neurological disorders may be white matterdisorders (a group of diseases that affects nerve fibers in the CNS)including but not limited to, Pelizaeus-Merzbacher disease,Hypomyelination with atrophy of basal ganglia and cerebellum,Aicardi-Goutières syndrome, Megalencephalic leukoencephalopathy withsubcortical cysts, Congenital muscular dystrophies, Myotonic dystrophy,Wilson disease, Lowe syndrome, Sjögren-Larsson syndrome, PIBD or Taysyndrome, Cockayne's disease, erebrotendinous xanthomatosis, Zellwegersyndrome, Neonatal adrenoleukodystrophy, Infantile Refsum disease,Zellweger-like syndrome, Pseudo-Zellweger syndrome, Pseudo-neonataladrenoleukodystrophy, Bifunctional protein deficiency, X-linkedadrenoleukodystrophy and adrenomyeloneuropathy and Refsum disease.

In some embodiments, the neurological disorders may be lysosomal storagedisorders (LSDs) caused by the inability of cells in the CNS to breakdown metabolic end products, including but not limited to Niemann-Pickdisease (a LSD resulting from inherited deficiency in acidsphingomyelinase (ASM); Metachromatic leukodystrophy (MLD) (a LSDcharacterized by accumulation of sulfatides in glial cells and neurons,the result of an inherited deficiency of arylsulfatase A (ARSA));Globoid-cell leukodystrophy (GLD) (a LSD caused by mutations ingalactosylceramidase); Fabry disease (a LSD caused by mutations in thealpha-galactosidase A (GLA) gene); Gaucher disease (caused by mutationsin the beta-glucocerebrosidase (GBA) gene); GM1/GM2 gangliosidosis;Mucopolysaccharidoses disorder; Pompe disease; and Neuronal ceroidlipofuscinosis.

In another embodiment, the neurological disease, disorder and/orcondition is Friedreich's Ataxia. In one embodiment, the AAV particleused to treat Friedreich's Ataxia comprises a nucleic acid sequence suchas, but not limited to, SEQ ID NO: 979 or a fragment or variant thereof,wherein the payload is replaced by Frataxin or any other payload knownin the art for treating Friedreich's Ataxia.

In another embodiment, the neurological disease, disorder and/orcondition is Amyotrophic lateral sclerosis (ALS). In one embodiment theAAV particle used to treat ALS comprises a nucleic acid sequence suchas, but not limited to, SEQ ID NO: 979 or a fragment or variant thereof,wherein the payload is replaced by replaced by an shRNA, miRNA, siRNA,RNAi for SOD1 or any other payload known in the art for treating ALS.

In another embodiment, the neurological disease, disorder and/orcondition is Huntington's disease. In one embodiment the AAV particleused to treat Huntington's disease comprises a nucleic acid sequencesuch as, but not limited to, SEQ ID NO: 979 or a fragment or variantthereof, wherein the payload is replaced by replaced by an shRNA, miRNA,siRNA, RNAi for Htt or any other payload known in the art for treatingHuntington's disease.

In another embodiment, the neurological disease, disorder or conditionis spinal muscular atrophy (SMA). In another embodiment, theneurological disease, disorder and/or condition is Friedreich's Ataxia.In one embodiment the AAV particle used to treat SMN comprises a nucleicacid sequence such as, but not limited to, SEQ ID NO: 979 or a fragmentor variant thereof, wherein the payload is replaced by Frataxin or anyother payload known in the art for treating SMA.

Parkinson's Disease

In one embodiment, the neurological disease, disorder and/or conditionis Parkinson's disease. In one embodiment the AAV particle used to treatParkinson's disease comprises a payload such as, but not limited to, SEQID NO: 979 or a fragment or variant thereof.

In one embodiment, the subject is a human patient who has a minimummotor score of about 30 to a maximum score of about 100, about 10 to amaximum score of about 100, about 20 to a maximum score of about 100 inthe Unified Parkinson's Disease Rating Scale.

In one embodiment, the subject has been diagnosed with Parkinson'sdisease within the past 5 years prior to treatment with the compositionsdescribed herein. As a non-limiting example, the subject may have beendiagnosed with Parkinson's disease within a week, 2 weeks, 3 weeks, 4weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 1 month, 2 months, 3 months,4 months, 5 months, 6

months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months,13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 1year, 2 years, 3 years, 4 years or less than 5 years prior to treatmentwith the compositions described herein.

In one embodiment, the subject has been diagnosed with Parkinson'sdisease between 5 and 10 years prior to treatment with the compositionsdescribed herein. As a non-limiting example, the subject may have beendiagnosed with Parkinson's disease 5, 5.5., 6, 6.5, 7, 7.5, 8, 8.5, 9,9.5 or 10 years prior to treatment with the compositions describedherein.

In one embodiment, the subject has been diagnosed with Parkinson'sdisease more than 10 years prior to treatment with the compositionsdescribed herein. As a non-limiting example, the subject may have beendiagnosed with Parkinson's disease 10.5, 11, 11.5, 12, 12.5, 13, 13.5,14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, 20, 20.5,21, 21.5, 22, 22.5, 23, 23.5, 24 or more than 24 years prior totreatment with the compositions described herein. In one embodiment, asubject is 50-65 years of age. As a non-limiting example, the subject is50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, or 65 yearsof age. As a non-limiting example, the subject is 50 years of age. As anon-limiting example, the subject is 51 years of age. As a non-limitingexample, the subject is 52 years of age. As a non-limiting example, thesubject is 53 years of age. As a non-limiting example, the subject is 54years of age. As a non-limiting example, the subject is 55 years of age.As a non-limiting example, the subject is 56 years of age. As anon-limiting example, the subject is 57 years of age. As a non-limitingexample, the subject is 58 years of age. As a non-limiting example, thesubject is 59 years of age. As a non-limiting example, the subject is 60years of age. As a non-limiting example, the subject is 61 years of age.As a non-limiting example, the subject is 62 years of age. As anon-limiting example, the subject is 63 years of age. As a non-limitingexample, the subject is 64 years of age. As a non-limiting example, thesubject is 65 years of age.

In one embodiment, a subject is 30 to 50 years of age. As a non-limitingexample, the subject is 30 years of age. As a non-limiting example, thesubject is 31 years of age. As a non-limiting example, the subject is 32years of age. As a non-limiting example, the subject is 33 years of age.As a non-limiting example, the subject is 34 years of age. As anon-limiting example, the subject is 35 years of age. As a non-limitingexample, the subject is 36 years of age. As a non-limiting example, thesubject is 37 years of age. As a non-limiting example, the subject is 38years of age. As a non-limiting example, the subject is 39 years of age.As a non-limiting example, the subject is 40 years of age. As anon-limiting example, the subject is 41 years of age. As a non-limitingexample, the subject is 42 years of age. As a non-limiting example, thesubject is 43 years of age. As a non-limiting example, the subject is 44years of age. As a non-limiting example, the subject is 45 years of age.As a non-limiting example, the subject is 46 years of age. As anon-limiting example, the subject is 47 years of age. As a non-limitingexample, the subject is 48 years of age. As a non-limiting example, thesubject is 49 years of age. As a non-limiting example, the subject is 50years of age.

In one embodiment, a subject is 65 to 85 years of age. As a non-limitingexample, the subject is 65 years of age. As a non-limiting example, thesubject is 66 years of age. As a non-limiting example, the subject is 67years of age. As a non-limiting example, the subject is 68 years of age.As a non-limiting example, the subject is 69 years of age. As anon-limiting example, the subject is 70 years of age. As a non-limitingexample, the subject is 71 years of age. As a non-limiting example, thesubject is 72 years of age. As a non-limiting example, the subject is 73years of age. As a non-limiting example, the subject is 74 years of age.As a non-limiting example, the subject is 75 years of age. As anon-limiting example, the subject is 76 years of age. As a non-limitingexample, the subject is 77 years of age. As a non-limiting example, thesubject is 78 years of age. As a non-limiting example, the subject is 79years of age. As a non-limiting example, the subject is 80 years of age.As a non-limiting example, the subject is 81 years of age. As anon-limiting example, the subject is 82 years of age. As a non-limitingexample, the subject is 83 years of age. As a non-limiting example, thesubject is 84 years of age. As a non-limiting example, the subject is 85years of age.

In one embodiment, a subject has seen a change in motor symptoms such astremors and movements prior to administration of the compositiondescribed herein. Non-limiting examples of tremors include, unilateralor bilateral mild tremors, bilateral or midline moderate tremors orintractable tremors. Non-limiting examples of movements include mildbradykinesia, moderate bradykinesia, severe bradykinesia and morningakinesia.

In one embodiment, a subject may have changes in balance such as, butnot limited to, impaired balance, impaired righting reflexes,significant balance disorder or falling.

In one embodiment, a subject may have a reduced quality of life. As anon-limiting example, the subject may have a moderate impact on theirquality of life such as experiencing some limitations to activities ofdaily living. As another non-limiting example, the subject may have aquality of life which has been diminished by illness.

In one embodiment, a subject has seen a change in non-motor symptomsprior to administration of the composition described herein. As anon-limiting example, the subject may have mild to moderate cognitiveimpairment prior to administration to the composition described herein.As another non-limiting example, the subject may have significantcognitive impairment such as dementia which may also include behavioraldisturbances such as hallucinations.

In one embodiment, a subject may have a satisfactory response withlimited fluctuations on one or more dopaminergic medications prior toadministration of the compositions described herein.

In one embodiment, a subject may have motor fluctuations causing mild tomoderate disability on one or more dopaminergic medications prior toadministration of the compositions described herein.

In one embodiment, a subject may have medically refractory motorfluctuations consisting of “wearing off” and/or levodopa-induceddyskinesias causing significant disability prior to administration ofthe compositions described herein.

In one embodiment, a subject may have mild symptoms associated withParkinson's disease such as, but not limited to, no cognitiveimpairment, diagnosed within the past 5 years, satisfactory responsewith limited fluctuations on one or more dopaminergic medications,unilateral or bilateral mild tremors, little to no impact on the qualityof life, and/or no balance impairment.

In one embodiment, a subject may have moderate symptoms associated withParkinson's disease such as, but not limited to, mild to moderatecognitive impairment, first signs of impaired balance and rightingreflexes, motor fluctuations causing mild-moderate disability on one ormore dopaminergic medications, diagnosed within the past 5 to 10 years,bilateral or midline moderate tremors, moderate bradykinesia and/orsubject experiencing some limitations to activities of daily living.

In one embodiment, a subject may have advanced symptoms associated withParkinson's disease such as, but not limited to, being diagnosed withParkinson's more than 10 years, medium refractory motor fluctuationswearing off and/or levodopa-induced dyskinesia causing significantdisability, intractable tremors, significant balance disorder and/orfalling, significant cognitive impairment (such as dementia with orwithout behavioral disturbances), sever bradykinesia, quality of lifemarkedly diminished by illness and/or morning akinesia.

In one embodiment, a subject has been referred to a movement disorderspecialist (MDS) but has not undergone deep brain stimulation.

In one embodiment, a subject is using DUOPA™ in combination with thecompositions described herein. As a non-limiting example, the subjectmay have success with using DUOPA™ alone. As a non-limiting example, thesubject may not have any success or limited success using DUOPA™ alone.

In one embodiment, a subject is one who was a candidate for surgicalintervention including, but not limited to, deep-brain stimulation. As anon-limiting example, deep-brain stimulation was suggested due todisabling motor complications despite treatment with optimalanti-Parkinsonian medication.

In one embodiment, a subject has an average on-time of 7.5-14 hoursbased on the subject diary. As a non-limiting example, the averageon-time is 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6,8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10, 10.1,10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3,11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12, 12.1, 12.2, 12.3, 12.4, 12.5,12.6, 12.7, 12.8, 12.9, 13, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7,13.8, 13.9, or 14 hours. In one embodiment, a subject has an averageon-time of 10.5 hours based on the subject diary.

In one embodiment, a subject experiences about 1.5 hours more of diaryon-time (without troublesome dyskinesia), as compared to baseline, 6months after administration of the present disclosure. As a non-limitingexample, a subject experiences about 1.5 hours more of diary on-time 6months after administration of the present disclosure at a dose volumeof up to 900 uL per putamen and a total dose of 4.5×10¹² vector genome.

In one embodiment, a subject experiences about 2.2 hours more of diaryon-time (without troublesome dyskinesia), as compared to baseline, 6months after administration of the present disclosure. As a non-limitingexample, a subject experiences about 2.2 hours more of diary on-time 6months after administration of the present disclosure at a dose volumeof up to 900 uL per putamen and a total dose of 1.5×10¹² vector genome.

In one embodiment, a subject experiences about 4 hours more of diaryon-time (without troublesome dyskinesia), as compared to baseline, 6months after administration of the present disclosure. As a non-limitingexample, a subject experiences about 4 hours more of diary on-time 6months after administration of the present disclosure at a dose volumeof up to 900 uL per putamen and a total dose of 1.5×10¹² vector genome.

In one embodiment, a subject experiences about 1.6 hours more of diaryon-time (without troublesome dyskinesia), as compared to baseline, 12months after administration of the present disclosure. As a non-limitingexample, a subject experiences about 1.6 hours more of diary on-time 12months after administration of the present disclosure at a dose volumeof up to 450 uL per putamen and a total dose of 7.5×10¹¹ vector genome.

In one embodiment, a subject experiences about 3.3 hours more of diaryon-time (without troublesome dyskinesia), as compared to baseline, 12months after administration of the present disclosure. As a non-limitingexample, a subject experiences about 3.3 hours more of diary on-time 12months after administration of the present disclosure at a dose volumeof up to 900 uL per putamen and a total dose of 1.5×10¹² vector genome.

In one embodiment, a subject experiences about 4 hours more of diaryon-time (without troublesome dyskinesia), as compared to baseline, 12months after administration of the present disclosure. As a non-limitingexample, a subject experiences about 4 hours more of diary on-time 12months after administration of the present disclosure at a dose volumeof up to 900 uL per putamen and a total dose of 1.5×10¹² vector genome.

In one embodiment, a subject experiences about 2.3 hours more of diaryon-time (without troublesome dyskinesia), as compared to baseline, 24months after administration of the present disclosure. As a non-limitingexample, a subject experiences about 2.3 hours more of diary on-time 24months after administration of the present disclosure at a dose volumeof up to 450 uL per putamen and a total dose of 7.5×10¹¹ vector genome.

In one embodiment, a subject has an average off-time of 2-7 hours basedon the subject diary. As a non-limiting example, the average off-time is2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4,3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9,5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4,6.5, 6.6, 6.7, 6.8, 6.9, or 7. In one embodiment, a subject has anaverage off-time of 4.6 hours based on the subject diary.

In one embodiment, a subject experiences about 1.3 hours less of diaryoff-time, as compared to baseline, 6 months after administration of thepresent disclosure. As a non-limiting example, a subject experiencesabout 1.3 hours less of diary off-time 6 months after administration ofthe present disclosure at a dose volume of up to 900 uL per putamen anda total dose of 4.5×10¹² vector genome.

In one embodiment, a subject experiences about 1.1 hours less of diaryoff-time, as compared to baseline, 6 months after administration of thepresent disclosure. As a non-limiting example, a subject experiencesabout 1.1 hours less of diary off-time 6 months after administration ofthe present disclosure at a dose volume of up to 900 uL per putamen anda total dose of 1.5×10¹² vector genome.

In one embodiment, a subject experiences about 0.8 hours less of diaryoff-time, as compared to baseline, 6 months after administration of thepresent disclosure. As a non-limiting example, a subject experiencesabout 0.8 hours less of diary off-time 6 months after administration ofthe present disclosure at a dose volume of up to 450 uL per putamen anda total dose of 7.5×10¹¹ vector genome.

In one embodiment, a subject experiences about 2.3 hours less of diaryoff-time, as compared to baseline, 12 months after administration of thepresent disclosure. As a non-limiting example, a subject experiencesabout 2.3 hours less of diary off-time 12 months after administration ofthe present disclosure at a dose volume of up to 900 uL per putamen anda total dose of 1.5×10¹² vector genome.

In one embodiment, a subject experiences about 1.4 hours less of diaryoff-time, as compared to baseline, 12 months after administration of thepresent disclosure. As a non-limiting example, a subject experiencesabout 1.4 hours less of diary off-time 12 months after administration ofthe present disclosure at a dose volume of up to 450 uL per putamen anda total dose of 7.5×10¹¹ vector genome.

In one embodiment, a subject experiences about 1.8 hours less of diaryoff-time, as compared to baseline, 24 months after administration of thepresent disclosure. As a non-limiting example, a subject experiencesabout 1.8 hours less of diary off-time 24 months after administration ofthe present disclosure at a dose volume of up to 450 uL per putamen anda total dose of 7.5×10¹¹ vector genome.

In one embodiment, a subject experiences 10% less diary off-time 6months after administration of the present disclosure. In oneembodiment, a subject experiences 10% less diary off-time 12 monthsafter administration of the present disclosure.

In one embodiment, a subject experiences 20% less diary off-time 6months after administration of the present disclosure. In oneembodiment, a subject experiences 20% less diary off-time 12 monthsafter administration of the present disclosure.

In one embodiment, a subject experiences 30% less diary off-time 6months after administration of the present disclosure. In oneembodiment, a subject experiences 30% less diary off-time 12 monthsafter administration of the present disclosure.

In one embodiment, a subject experiences 40% less diary off-time 6months after administration of the present disclosure. In oneembodiment, a subject experiences 40% less diary off-time 12 monthsafter administration of the present disclosure.

In one embodiment, a subject experiences 50% less diary off-time 6months after administration of the present disclosure. In oneembodiment, a subject experiences 50% less diary off-time 12 monthsafter administration of the present disclosure.

In one embodiment, a subject experiences 60% less diary off-time 6months after administration of the present disclosure. In oneembodiment, a subject experiences 60% less diary off-time 12 monthsafter administration of the present disclosure.

In one embodiment, a subject experiences 70% less diary off-time 6months after administration of the present disclosure. In oneembodiment, a subject experiences 70% less diary off-time 12 monthsafter administration of the present disclosure.

In one embodiment, a subject experiences 80% less diary off-time 6months after administration of the present disclosure. In oneembodiment, a subject experiences 80% less diary off-time 12 monthsafter administration of the present disclosure.

In one embodiment, a subject experiences 90% less diary off-time 6months after administration of the present disclosure. In oneembodiment, a subject experiences 90% less diary off-time 12 monthsafter administration of the present disclosure.

In one embodiment, a subject experiences 95% less diary off-time 6months after administration of the present disclosure. In oneembodiment, a subject experiences 95% less diary off-time 12 monthsafter administration of the present disclosure.

In one embodiment, a subject's UPDRS-3 (or UPDRS-III) medication scoreis evaluated prior to administration of the present disclosure. As anon-limiting example, the subject's UPDRS-3 (or UPDRS-III) medicationscore prior to administration of the present disclosure is between 1-40,1-10, 1-15, 1-20, 1-25, 1-30, 1-35, 1-40, 5-10, 5-15, 5-20, 5-25, 5-30,5-35, 5-40, 10-15, 10-20, 10-25, 10-30, 10-35, 10-40, 15-20, 15-25,15-30, 15-35, 15-40, 20-25, 20-30, 20-35, 20-40, 25-30, 25-35, 25-40,30-35, 30-40, or 35-40. As a non-limiting example, the subject's UPDRS-3(or UPDRS-III) medication score prior to administration of the presentdisclosure is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,36, 37, 38, 39, or 40. As a non-limiting example, the subject's UPDRS-3(or UPDRS-III) medication score prior to administration of the presentdisclosure is 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11,11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12, 12.1, 12.2,12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13, 13.1, 13.2, 13.3, 13.4,13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6,14.7, 14.8, 14.9, or 15. As a non-limiting example, the subjects UPDRS-3medication score prior to administration of the present disclosure is13.5.

In one embodiment, a subject's UPDRS-3 (or UPDRS-III) medication scoreis reduced after administration of the present disclosure.

A subject's UPDRS-3 (or UPDRS-III) medication score may be reduced by apercentage such as, but not limited to, 10%, 15%, 20%, 25%, 30%, 35%,40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more than95%. As a non-limiting example, a subject's UPDRS-3 score is reduced 10%after administration of the present disclosure. As a non-limitingexample, a subject's UPDRS-3 score is reduced 20% after administrationof the present disclosure. As a non-limiting example, a subject'sUPDRS-3 score is reduced 30% after administration of the presentdisclosure. As a non-limiting example, a subject's UPDRS-3 score isreduced 40% after administration of the present disclosure. As anon-limiting example, a subject's UPDRS-3 score is reduced 50% afteradministration of the present disclosure. As a non-limiting example, asubject's UPDRS-3 score is reduced 60% after administration of thepresent disclosure. As a non-limiting example, a subject's UPDRS-3 scoreis reduced 70% after administration of the present disclosure. As anon-limiting example, a subject's UPDRS-3 score is reduced 80% afteradministration of the present disclosure. As a non-limiting example, asubject's UPDRS-3 score is reduced 90% after administration of thepresent disclosure.

A subject's UPDRS-3 (or UPDRS-III) medication score may change by 0.1,0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6,1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1,3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6,4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1,6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6,7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1,9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4,10.5, 10.6, 10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6,11.7, 11.8, 11.9, 12, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8,12.9, 13, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14,14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or 15 points. As anon-limiting example, a subject's UPDRS-3 score is changed by 0.4points. As a non-limiting example, a subject's UPDRS-3 score is changedby 1.6 points. As a non-limiting example, a subject's UPDRS-3 score ischanged by 1.8 points. As a non-limiting example, a subject's UPDRS-3score is changed by 8.6 points. As a non-limiting example, a subject'sUPDRS-3 score is changed by 9.6 points.

A subject's UPDRS-3 (or UPDRS-III) medication score may increase by 0.1,0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6,1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1,3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6,4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1,6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6,7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1,9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4,10.5, 10.6, 10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6,11.7, 11.8, 11.9, 12, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8,12.9, 13, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14,14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or 15 points. As anon-limiting example, a subject's UPDRS-3 score is increased by 0.4points. As a non-limiting example, a subject's UPDRS-3 score isincreased by 1.6 points. As a non-limiting example, a subject's UPDRS-3score is increased by 1.8 points. As a non-limiting example, a subject'sUPDRS-3 score is increased by 8.6 points. As a non-limiting example, asubject's UPDRS-3 score is increased by 9.6 points.

A subject's UPDRS-3 (or UPDRS-III) medication score may decrease by 0.1,0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6,1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1,3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6,4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1,6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6,7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1,9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4,10.5, 10.6, 10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6,11.7, 11.8, 11.9, 12, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8,12.9, 13, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14,14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or 15 points. As anon-limiting example, a subject's UPDRS-3 score is decreased by 0.4points. As a non-limiting example, a subject's UPDRS-3 score isdecreased by 1.6 points. As a non-limiting example, a subject's UPDRS-3score is decreased by 1.8 points. As a non-limiting example, a subject'sUPDRS-3 score is decrease by 8.6 points. As a non-limiting example, asubject's UPDRS-3 score is decreased by 9.6 points.

In one embodiment, a subject's UPDRS-3 (or UPDRS-III) medication scoreis reduced by 8.6 at 6 months after administration of the presentdisclosure at a dose volume of up to 900 uL per putamen and a total doseof 4.5×10¹² vector genome.

In one embodiment, a subject's UPDRS-3 (or UPDRS-III) medication scoreis reduced by 9.6 at 6 months after administration of the presentdisclosure at a dose volume of up to 900 uL per putamen and a total doseof 1.5×10¹² vector genome.

In one embodiment, a subject's UPDRS-3 (or UPDRS-III) medication scoreis reduced by 9.6 at 12 months after administration of the presentdisclosure at a dose volume of up to 900 uL per putamen and a total doseof 1.5×10¹² vector genome.

In one embodiment, a subject's average amount of Parkinson's diseasemedication was about 1500 mg per day prior to administration of thepresent disclosure. As a non-limiting example, the Parkinson's diseasemedication is levodopa.

In one embodiment, a subject's UPDRS-II score is evaluated prior toadministration of the present disclosure. The UPDRS-II score of asubject prior to administration of the present disclosure is between 20and 50, such as, but not limited to, 20, 21, 22, 23, 24, 25, 26, 27, 28,29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,47, 48, 49, and 50.

A subject's UPDRS-2 (or UPDRS-II) score may change by 0.1, 0.2, 0.3,0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8,1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3,3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8,4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3,6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8,7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3,9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6,10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8,11.9, 12, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13,13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1, 14.2,14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or 15 points.

A subject's UPDRS-2 (or UPDRS-II) score may increase by 0.1, 0.2, 0.3,0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8,1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3,3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8,4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3,6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8,7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3,9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6,10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8,11.9, 12, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13,13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1, 14.2,14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or 15 points.

A subject's UPDRS-2 (or UPDRS-II) score may decrease by 0.1, 0.2, 0.3,0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8,1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3,3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8,4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3,6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8,7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3,9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6,10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8,11.9, 12, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13,13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1, 14.2,14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or 15 points. As anon-limiting example, a 3.6 point reduction is seen 12 months afteradministration with the present disclosure. As a non-limiting example, a3.6 point reduction is seen 6 months after administration with thepresent disclosure.

In one embodiment, a subject's UPDRS-II score is decreased by 2 to 4points after administration of the present disclosure as compared to theUPDRS-II score prior to administration.

In one embodiment, a subject's UPDRS-II score is decreased by 2 to 3points 6 months after administration of the present disclosure at a dosevolume of up to 900 uL per putamen and a total dose of 1.5×10¹² vectorgenome, as compared to the UPDRS-II medication score prior toadministration.

In one embodiment, a subject's UPDRS-II score is decreased by 2 to 3points 12 months after administration of the present disclosure at adose volume of up to 900 uL per putamen and a total dose of 1.5×10¹²vector genome, as compared to the UPDRS-II medication score prior toadministration.

In one embodiment, a subject's UPDRS-II score is decreased by 3 to 4points 6 months after administration of the present disclosure at a dosevolume of up to 900 uL per putamen and a total dose of 4.5×10¹² vectorgenome, as compared to the UPDRS-II medication score prior toadministration.

In one embodiment, the present disclosure is used to improve a subject'smotor function.

In one embodiment, the present disclosure is used to control a subject'smotor function and improve their quality of life.

In one embodiment, the present disclosure is used to reduce the dosageof Parkinson's medication a subject needs to take to improve a subject'smotor function.

In one embodiment, a single administration of the present disclosureinto the putamen of a subject provides improved motor function ascompared to motor function prior to treatment.

In one embodiment, a single administration of the present disclosure into the putamen of a subject provides improved motor function and areduction in the amount of levodopa the subject requires to managesymptoms.

In one embodiment, a single administration of the present disclosure into the putamen of a subject provides improved motor function and areduction in the amount of dopaminergic medication the subject requiresto manage symptoms.

In one embodiment, the amount the daily dose of Parkinson's medication(e.g., Levodopa) is reduced by 10-50% after administration of thepresent disclosure. As a non-limiting example, the reduction is seen 6months after administration of the present disclosure. As a non-limitingexample, the reduction is seen 12 months after administration of thepresent disclosure.

In one embodiment, the amount the daily dose of Parkinson's medication(e.g., Levodopa) is reduced by 10-20% after administration of thepresent disclosure. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 10%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is11%. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 12%. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 13%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is14%. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 15%. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 16%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is17%. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 18%. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 19%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is20%.

In one embodiment, a 14% reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 6 months after administration of the present disclosureat a dose volume of up to 450 uL per putamen and a total dose of7.5×10¹¹ vector genome.

In one embodiment, a 10% reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 12 months after administration of the present disclosureat a dose volume of up to 450 uL per putamen and a total dose of7.5×10¹¹ vector genome.

In one embodiment, the amount the daily dose of Parkinson's medication(e.g., Levodopa) is reduced by 20-30% after administration of thepresent disclosure. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 20%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is21%. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 22%. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 23%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is24%. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 25%. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 26%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is27%. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 28%. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 29%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is30%.

In one embodiment, a 27% reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 12 months after administration of the present disclosureat a dose volume of up to 900 uL per putamen and a total dose of1.5×10¹² vector genome.

In one embodiment, a 28% reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 12 months after administration of the present disclosureat a dose volume of up to 900 uL per putamen and a total dose of1.5×10¹² vector genome.

In one embodiment, a 29% reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 12 months after administration of the present disclosureat a dose volume of up to 900 uL per putamen and a total dose of1.5×10¹² vector genome.

In one embodiment, the amount the daily dose of Parkinson's medication(e.g., Levodopa) is reduced by 30-40% after administration of thepresent disclosure. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 30%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is31%. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 32%. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 33%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is34%. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 35%. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 36%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is37%. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 38%. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 39%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is40%.

In one embodiment, a 34% reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 6 months after administration of the present disclosureat a dose volume of up to 900 uL per putamen and a total dose of1.5×10¹² vector genome.

In one embodiment, the amount the daily dose of Parkinson's medication(e.g., Levodopa) is reduced by 40-50% after administration of thepresent disclosure. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 40%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is41%. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 42%. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 43%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is44%. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 45%. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 46%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is47%. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 48%. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 49%. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is50%.

In one embodiment, a 34% reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 6 months after administration of the present disclosureat a dose volume of up to 900 uL per putamen and a total dose of4.5×10¹² vector genome.

In one embodiment, the amount the daily dose of Parkinson's medication(e.g., Levodopa) is reduced by 108-641 mg after administration of thepresent disclosure. As a non-limiting example, the reduction is seen 6months after administration of the present disclosure. As a non-limitingexample, the reduction is seen 12 months after administration of thepresent disclosure.

In one embodiment, the amount the daily dose of Parkinson's medication(e.g., Levodopa) is reduced by 108-339 mg after administration of thepresent disclosure. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 108 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 134mg. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 159 mg. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 154 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 208mg. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 231 mg. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 254 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 276mg. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 298 mg. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 319 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 339mg.

In one embodiment, a 208 mg reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 6 months after administration of the present disclosureat a dose volume of up to 450 uL per putamen and a total dose of7.5×10¹¹ vector genome.

In one embodiment, a 108 mg reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 12 months after administration of the present disclosureat a dose volume of up to 450 uL per putamen and a total dose of7.5×10¹¹ vector genome.

In one embodiment, the amount the daily dose of Parkinson's medication(e.g., Levodopa) is reduced by 339-505 mg after administration of thepresent disclosure. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 339 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 358mg. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 377 mg. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 396 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 413mg. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 430 mg. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 446 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 462mg. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 477 mg. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 491 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 505mg.

In one embodiment, a 462 mg reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 12 months after administration of the present disclosureat a dose volume of up to 900 uL per putamen and a total dose of1.5×10¹² vector genome.

In one embodiment, a 477 mg reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 12 months after administration of the present disclosureat a dose volume of up to 900 uL per putamen and a total dose of1.5×10¹² vector genome.

In one embodiment, a 491 mg reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 12 months after administration of the present disclosureat a dose volume of up to 900 uL per putamen and a total dose of1.5×10¹² vector genome.

In one embodiment, the amount the daily dose of Parkinson's medication(e.g., Levodopa) is reduced by 505-606 mg after administration of thepresent disclosure. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 505 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 518mg. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 530 mg. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 542 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 553mg. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 563 mg. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 573 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 582mg. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 591 mg. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 599 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 606mg.

In one embodiment, a 553 mg reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 6 months after administration of the present disclosureat a dose volume of up to 900 uL per putamen and a total dose of1.5×10¹² vector genome.

In one embodiment, the amount the daily dose of Parkinson's medication(e.g., Levodopa) is reduced by 606-641 mg after administration of thepresent disclosure. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 606 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 612mg. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 618 mg. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 623 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 628mg. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 632 mg. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 635 mg. As a non-limitingexample, the reduction of Parkinson's medication (e.g., Levodopa) is 637mg. As a non-limiting example, the reduction of Parkinson's medication(e.g., Levodopa) is 639 mg. As a non-limiting example, the reduction ofParkinson's medication (e.g., Levodopa) is 641 mg.

In one embodiment, a 553 mg reduction in the amount of the daily dose ofParkinson's medication (e.g., Levodopa) required by a subject to managesymptoms occurs 6 months after administration of the present disclosureat a dose volume of up to 900 uL per putamen and a total dose of4.5×10¹² vector genome.

In one embodiment, the putaminal AADC enzyme activity is increased in asubject after administration with the present disclosure. As anon-limiting example, the increase is seen for at least 6 monthsrelative to the baseline.

In one embodiment, the putaminal AADC enzyme activity is increased by10-20% in a subject after administration of the present disclosure. As anon-limiting example, the increase in putaminal AADC enzyme activity isabout 10%. As a non-limiting example, the increase in putaminal AADCenzyme activity is about 11%. As a non-limiting example, the increase inputaminal AADC enzyme activity is about 12%. As a non-limiting example,the increase in putaminal AADC enzyme activity is about 13%. As anon-limiting example, the increase in putaminal AADC enzyme activity isabout 14%. As a non-limiting example, the increase in putaminal AADCenzyme activity is about 15%. As a non-limiting example, the increase inputaminal AADC enzyme activity is about 16%. As a non-limiting example,the increase in putaminal AADC enzyme activity is about 17%. As anon-limiting example, the increase in putaminal AADC enzyme activity isabout 18%. As a non-limiting example, the increase in putaminal AADCenzyme activity is about 19%. As a non-limiting example, the increase inputaminal AADC enzyme activity is about 20%.

In one embodiment, the putaminal AADC enzyme activity is increased byabout 13% in a subject after administration of the present disclosure ata dose volume of up to 450 uL per putamen and a total dose of 7.5×10¹¹vector genome.

In one embodiment, the putaminal AADC enzyme activity is increased by50-60% in a subject after administration of the present disclosure. As anon-limiting example, the increase in putaminal AADC enzyme activity isabout 50%. As a non-limiting example, the increase in putaminal AADCenzyme activity is about 51%. As a non-limiting example, the increase inputaminal AADC enzyme activity is about 52%. As a non-limiting example,the increase in putaminal AADC enzyme activity is about 53%. As anon-limiting example, the increase in putaminal AADC enzyme activity isabout 54%. As a non-limiting example, the increase in putaminal AADCenzyme activity is about 55%. As a non-limiting example, the increase inputaminal AADC enzyme activity is about 56%. As a non-limiting example,the increase in putaminal AADC enzyme activity is about 57%. As anon-limiting example, the increase in putaminal AADC enzyme activity isabout 58%. As a non-limiting example, the increase in putaminal AADCenzyme activity is about 59%. As a non-limiting example, the increase inputaminal AADC enzyme activity is about 60%.

In one embodiment, the putaminal AADC enzyme activity is increased byabout 56% in a subject after administration of the present disclosure ata dose volume of up to 900 uL per putamen and a total dose of 1.5×10¹²vector genome.

In one embodiment, the putaminal AADC enzyme activity is increased by70-85% in a subject after administration of the present disclosure. As anon-limiting example, the increase in putaminal AADC enzyme activity isabout 70%. As a non-limiting example, the increase in putaminal AADCenzyme activity is about 71%. As a non-limiting example, the increase inputaminal AADC enzyme activity is about 72%. As a non-limiting example,the increase in putaminal AADC enzyme activity is about 73%. As anon-limiting example, the increase in putaminal AADC enzyme activity isabout 74%. As a non-limiting example, the increase in putaminal AADCenzyme activity is about 75%. As a non-limiting example, the increase inputaminal AADC enzyme activity is about 76%. As a non-limiting example,the increase in putaminal AADC enzyme activity is about 77%. As anon-limiting example, the increase in putaminal AADC enzyme activity isabout 78%. As a non-limiting example, the increase in putaminal AADCenzyme activity is about 79%. As a non-limiting example, the increase inputaminal AADC enzyme activity is about 80%. As a non-limiting example,the increase in putaminal AADC enzyme activity is about 81%. As anon-limiting example, the increase in putaminal AADC enzyme activity isabout 82%. As a non-limiting example, the increase in putaminal AADCenzyme activity is about 83%. As a non-limiting example, the increase inputaminal AADC enzyme activity is about 84%. As a non-limiting example,the increase in putaminal AADC enzyme activity is about 85%.

In one embodiment, the putaminal AADC enzyme activity is increased byabout 79% in a subject after administration of the present disclosure ata dose volume of up to 900 uL per putamen and a total dose of 4.5×10¹²vector genome.

In one embodiment, the dopamine level of a subject increased afteradministration of the present disclosure. As a non-limiting example, theamount of dopamine increased by 10-20%, 15-25%, 20-30%, 25-35%, 30-40%,35-45%, 40-50%, 45-55%, 50-60%, 55-65%, 60-70%, 65-75%, 70-80%, 75-85%,80-90%, 85-95%, 90-100%, or 95-100%.

Circadian Rhythm and Sleep-Wake Cycles

Circadian rhythms are physical, mental and behavioral changes that tendto follow a 24 hour cycle. Circadian rhythms can influence sleep-wakecycles, hormone release, body temperature and other bodily functions.Changes in the circadian rhythm can cause conditions and/or disordersuch as, but not limited to sleep disorders (e.g., insomnia),depression, bipolar disorder, seasonal affective disorder, obesity anddiabetes.

In one embodiment, the AAV particles described herein may be used totreat insomnia.

The sleep-wake cycle comprises periods of sleep and periods of wake.Generally, in a 24 hour period the total hours of sleep are less thanthe total hours of wakefulness. As a non-limiting example, thesleep-wake cycle comprises 7-9 hours of sleep and 15-17 hours ofwakefulness. As a non-limiting example, the sleep-wake cycle comprises 8hours of sleep and 16 hours of wakefulness. As a non-limiting example,the sleep-wake cycle comprises 8-10 hours of sleep and 14-16 hours ofwakefulness.

In one embodiment, the sleep-wake cycle of a subject is improved byadministering to the subject the AAV particles described herein.

In one embodiment, the sleep-wake cycle of a subject is regulated byadministering to the subject the AAV particles described herein. As anon-limiting example, the regulation may be the correction of moreperiods of sleep occurring at night and less periods of sleep occurring

In one embodiment, the sleep-wake cycle of a subject administered theAAV particles described herein improves as compared to the sleep-wakecycle of the subject prior to administration of the AAV particles. As anon-limiting example, the subject has an increased period of sleep and adecreased period of wakefulness. As another non-limiting example, thesubject has a decreased period of sleep and an increased period ofwakefulness.

In one embodiment, the sleep-wake cycle of a subject administered theAAV particles described herein is regulated as compared to thesleep-wake cycle of the subject prior to administration of the AAVparticles. As a non-limiting example, the length of the periods of sleepand the periods of wakefulness may be about the same (e.g., +/−1 hour)for at least 2 days. As another non-limiting example, the length of theperiods of sleep and the periods of wakefulness if a 24 hours period maybe within 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 1 hour, 1.5hours, or 2 hours of the previous 24 hour period.

In one embodiment, the amount of rapid eye movement (REM) sleep asubject experiences in a 24 hour period is altered after the subject isadministered the AAV particles described herein. REM sleep is generallyconsidered an active period of sleep marked by intense brain activitywhere brain waves are fast and desynchronized. An adult, on average,spends about 20-25% of their total daily sleep period in REM sleep. As anon-limiting example, the amount of REM sleep is decreased by 1%, 2%,3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%,35%, 40%, 45%, 50%, 55%, 60%, 65% or more than 65%. As a non-limitingexample, the amount of REM sleep is decreased by 1-10%, 5-10%, 5-15%,10-15%, 15-20%, 15-25%, 20-25%, 20-30%, 25-30%, 25-35%, 30-35%, 30-40%,35-40%, 40-50% or 40-60%. As a non-limiting example, the amount of REMsleep is increased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%,13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or morethan 65%. As a non-limiting example, the amount of REM sleep isincreased by 1-5%, 1-10%, 5-10%, 5-15%, 10-15%, 15-20%, 15-25%, 20-25%,20-30%, 25-30%, 25-35%, 30-35%, 30-40%, 35-40%, 40-50% or 40-60%.

In one embodiment, the amount of non-REM (NREM) sleep a subjectexperiences in a 24 hour period is altered after the subject isadministered the AAV particles described herein. NREM sleep is generallycharacterized by a reduction in physiological activity since as thebrain waves, measured by EEG, get slower and have greater amplitude.NREM has four stages: Stage 1 is the time of drowsiness or transitionfrom being awake to falling asleep where the brain waves and muscleactivity begin to slow; Stage 2 is a period of light sleep during whicheye movements stop and brain waves become slower with occasional burstsof rapid waves (sometimes called sleep spindles); Stage 3 and Stage 4(collectively referred to as slow wave sleep) are characterized by thepresence of slow brain waves (delta waves) interspersed with smallerfaster waves where there are no eye movements. An adult, on average,spends about 75-80% of their total daily sleep period in NREM sleep withabout half of their total daily sleep time in NREM stage 2 sleep.

In one embodiment, the amount of NREM sleep a subject experiences isaltered after the subject is administered the AAV particles describedherein. As a non-limiting example, the amount of NREM sleep is decreasedby 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%,20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or more than 65%. As anon-limiting example, the amount of NREM sleep is decreased by 1-10%,5-10%, 5-15%, 10-15%, 15-20%, 15-25%, 20-25%, 20-30%, 25-30%, 25-35%,30-35%, 30-40%, 35-40%, 40-50% or 40-60%. As a non-limiting example, theamount of NREM sleep is increased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%,10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65% or more than 65%. As a non-limiting example, the amount of NREMsleep is increased by 1-5%, 1-10%, 5-10%, 5-15%, 10-15%, 15-20%, 15-25%,20-25%, 20-30%, 25-30%, 25-35%, 30-35%, 30-40%, 35-40%, 40-50% or40-60%.

In one embodiment, the amount of NREM Stage 1 sleep a subjectexperiences is altered after the subject is administered the AAVparticles described herein. As a non-limiting example, the amount ofNREM Stage 1 sleep is decreased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%,10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65% or more than 65%. As a non-limiting example, the amount of NREMStage 1 sleep is decreased by 1-10%, 5-10%, 5-15%, 10-15%, 15-20%,15-25%, 20-25%, 20-30%, 25-30%, 25-35%, 30-35%, 30-40%, 35-40%, 40-50%or 40-60%. As a non-limiting example, the amount of NREM Stage 1 sleepis increased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%,14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or more than65%. As a non-limiting example, the amount of NREM Stage 1 sleep isincreased by 1-5%, 1-10%, 5-10%, 5-15%, 10-15%, 15-20%, 15-25%, 20-25%,20-30%, 25-30%, 25-35%, 30-35%, 30-40%, 35-40%, 40-50% or 40-60%.

In one embodiment, the amount of NREM Stage 2 sleep a subjectexperiences is altered after the subject is administered the AAVparticles described herein. As a non-limiting example, the amount ofNREM Stage 2 sleep is decreased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%,10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65% or more than 65%. As a non-limiting example, the amount of NREMStage 2 sleep is decreased by 1-10%, 5-10%, 5-15%, 10-15%, 15-20%,15-25%, 20-25%, 20-30%, 25-30%, 25-35%, 30-35%, 30-40%, 35-40%, 40-50%or 40-60%. As a non-limiting example, the amount of NREM Stage 2 sleepis increased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%,14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or more than65%. As a non-limiting example, the amount of NREM Stage 2 sleep isincreased by 1-5%, 1-10%, 5-10%, 5-15%, 10-15%, 15-20%, 15-25%, 20-25%,20-30%, 25-30%, 25-35%, 30-35%, 30-40%, 35-40%, 40-50% or 40-60%.

In one embodiment, the amount of NREM Stage 3 and 4 sleep a subjectexperiences is altered after the subject is administered the AAVparticles described herein. As a non-limiting example, the amount ofNREM Stage 3 and 4 sleep is decreased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%,9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%,55%, 60%, 65% or more than 65%. As a non-limiting example, the amount ofNREM Stage 3 and 4 sleep is decreased by 1-10%, 5-10%, 5-15%, 10-15%,15-20%, 15-25%, 20-25%, 20-30%, 25-30%, 25-35%, 30-35%, 30-40%, 35-40%,40-50% or 40-60%. As a non-limiting example, the amount of NREM Stage 3and 4 sleep is increased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%,11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65% or more than 65%. As a non-limiting example, the amount of NREMStage 3 and 4 sleep is increased by 1-5%, 1-10%, 5-10%, 5-15%, 10-15%,15-20%, 15-25%, 20-25%, 20-30%, 25-30%, 25-35%, 30-35%, 30-40%, 35-40%,40-50% or 40-60%.

In one embodiment, periods of NREM and REM cycles are more consistent ina subject after the subject is administered the AAV particles describedherein. Generally NREM and REM cycles alternate every 90 to 110 minutesfour to six times per night.

V. Kits and Devices Kits

In one embodiment, the disclosure provides a variety of kits forconveniently and/or effectively carrying out methods of the presentdisclosure. Typically kits will comprise sufficient amounts and/ornumbers of components to allow a user to perform multiple treatments ofa subject(s) and/or to perform multiple experiments.

Any of the AAV particles of the present disclosure may be comprised in akit. In some embodiments, kits may further include reagents and/orinstructions for creating and/or synthesizing compounds and/orcompositions of the present disclosure. In some embodiments, kits mayalso include one or more buffers. In some embodiments, kits of thedisclosure may include components for making protein or nucleic acidarrays or libraries and thus, may include, for example, solid supports.

In some embodiments, kit components may be packaged either in aqueousmedia or in lyophilized form. The container means of the kits willgenerally include at least one vial, test tube, flask, bottle, syringeor other container means, into which a component may be placed, andpreferably, suitably aliquoted. Where there is more than one kitcomponent, (labeling reagent and label may be packaged together), kitsmay also generally contain second, third or other additional containersinto which additional components may be separately placed. In someembodiments, kits may also comprise second container means forcontaining sterile, pharmaceutically acceptable buffers and/or otherdiluents. In some embodiments, various combinations of components may becomprised in one or more vial. Kits of the present disclosure may alsotypically include means for containing compounds and/or compositions ofthe present disclosure, e.g., proteins, nucleic acids, and any otherreagent containers in close confinement for commercial sale. Suchcontainers may include injection or blow-molded plastic containers intowhich desired vials are retained.

In some embodiments, kit components are provided in one and/or moreliquid solutions. In some embodiments, liquid solutions are aqueoussolutions, with sterile aqueous solutions being particularly preferred.In some embodiments, kit components may be provided as dried powder(s).When reagents and/or components are provided as dry powders, suchpowders may be reconstituted by the addition of suitable volumes ofsolvent. In some embodiments, it is envisioned that solvents may also beprovided in another container means. In some embodiments, labeling dyesare provided as dried powders. In some embodiments, it is contemplatedthat 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 120, 130, 140, 150,160, 170, 180, 190, 200, 300, 400, 500, 600, 700, 800, 900, 1000micrograms or at least or at most those amounts of dried dye areprovided in kits of the disclosure. In such embodiments, dye may then beresuspended in any suitable solvent, such as DMSO.

In some embodiments, kits may include instructions for employing kitcomponents as well the use of any other reagent not included in the kit.Instructions may include variations that may be implemented.

Devices

In some embodiments, AAV particles of the present disclosure may becombined with, coated onto or embedded in a device. Devices may include,but are not limited to stents, pumps, and/or other implantabletherapeutic device. Additionally AAV particles may be delivered to asubject while the subject is using a compression device such as, but notlimited to, a compression device to reduce the chances of deep veinthrombosis (DVT) in a subject.

The present disclosure provides for devices which may incorporate AAVparticles. These devices contain in a stable formulation the AAVparticles which may be immediately delivered to a subject in needthereof, such as a human patient.

Devices for administration may be employed to deliver the AAV particlesof the present disclosure according to single, multi- or split-dosingregimens taught herein.

Method and devices known in the art for multi-administration to cells,organs and tissues are contemplated for use in conjunction with themethods and compositions disclosed herein as embodiments of the presentdisclosure. These include, for example, those methods and devices havingmultiple needles, hybrid devices employing for example lumens orcatheters as well as devices utilizing heat, electric current orradiation driven mechanisms.

In some embodiments, AAV particles of the present disclosure may bedelivered using a device such as, but not limited to, a stent, a tube, acatheter, a pipe, a straw, needle and/or a duct. Methods of using thesedevices are described herein and are known in the art.

In one embodiment, the AAV particles of the present disclosure may beadministered to a subject using delivery systems which integrate imageguided therapy and integrate imaging such as, but not limited to, laser,MRgFUS, endoscopic and robotic surgery devices.

In one embodiment, the AAV particles of the present disclosure may beadministered to a subject using the CLEARPOINT® neuro interventionsystem by Mill Interventions, Inc. The CLEARPOINT® neuro interventionsystem may be used alone or in combination with any of the otheradministration methods and devices described herein. The CLEARPOINT®neuro intervention system helps to provide stereotactic guidance in theplacement and operation of instruments or devices during the planningand operation of neurological procedures.

In one embodiment, the AAV particles of the present disclosure may beadministered to a subject using the NEUROMATE® stereotactic robot systemby Renishaw PLC. The NEUROMATE® system may be used alone or incombination with any of the other administration methods and devicesdescribed herein. As a non-limiting example, the NEUROMATE® system maybe used with head holders, CT image localizers, frame attachments,remote controls and software.

In one embodiment, the AAV particles of the present disclosure may beadministered to a subject using the Elekta MICRODRIVE™ device by ElektaAB. The MICRODRIVE™ device may be used alone or in combination with anyof the other administration methods and devices described herein. As anon-limiting example, the MICRODRIVE™ device may be used to positionelectrodes (e.g., for micro electrode recording (MER), macro stimulationand deep brain stimulation (DBS) electrode implantation), implantationof catheters, tubes or DBS electrodes using cross-hair and A-P holdersto verify position, biopsies, injections and aspirations, brainlesioning, endoscope guidance and GAMMA KNIFE® radiosurgery.

In one embodiment, the AAV particles of the present disclosure may beadministered to a subject using the AXIIIS® stereotactic miniframe byMONTERIS® Medical, Inc. The AXIIIS® stereotactic miniframe may be usedalone or in combination with any of the other administration methods anddevices described herein. The AXIIIS® stereotactic miniframe is atrajectory alignment device which may be used for laser coagulation,biopsies, catheter placement, electrode implant, endoscopy, and clotevacuation. The miniframe allows for 360 degree interface and providesaccess to multiple intracranial targets with a simple adjustment.Further, the miniframe is compatible with MM.

In one embodiment, the AAV particles of the present disclosure may beadministered to a subject using the INTEGRA™ CRW® system by IntegraLifeSciences Corporation. The INTEGRA™ CRW® system may be used alone orin combination with any of the other administration methods and devicesdescribed herein. The CRW® system may be used for various applicationssuch as, but not limited to, stereotactic surgery, microsurgery,catheterization and biopsy. The CRW® system is designed to provideaccuracy to those who use the system (e.g., thumb lock screws, Vernierscaling, double bolt fixation, and a solid frame).

In one embodiment, the AAV particles of the present disclosure may beadministered to a subject using the EPOCH® solution system byStereotaxis, Inc. which may include the NIOBE® ES magnetic navigationsystem, the VDRIVE® robotic navigation system and/or the ODYSSEY®information solution (all by Stereotaxis, Inc.). The EPOCH® solutionsystem may be used alone or in combination with any of the otheradministration methods and devices described herein. As a non-limitingexample, the NIOBE® ES magnetic navigation system may be used toaccurately contact a subject. As another non-limiting example the NIOBE®ES magnetic system may be used with the VDRIVE® robotic navigationsystem to provide precise movement and stability.

In one embodiment, the AAV particles of the present disclosure may beadministered to a subject using a NeuroStation workstation which usesframeless stereotactic methods to provide image-guidance forapplications such as, but not limited to, surgical planning, biopsies,craniotomies, endoscopy, intra-operative ultrasound and radiationtherapy.

In one embodiment, the AAV particles of the present disclosure may beadministered to a subject using a robotic stereotaxis system such as,but not limited to the device described in U.S. Pat. No. 5,078,140, thecontents of which are herein incorporated by reference in its entirety.The robotic arm of the device may be used to precisely orient thesurgical tools or other implements used to conduct a procedure.

In one embodiment, the AAV particles of the present disclosure may beadministered to a subject using an automatic delivery system such as,but not limited to the device described in U.S. Pat. No. 5,865,744, thecontents of which are herein incorporated by reference in its entirety.Based on the images gathered by the delivery system, the computeradjusts the administration of the needle to be the appropriate depth forthe particular subject.

In one embodiment, the AAV particles of the present disclosure may beadministered to a subject who is simultaneously using duringadministration, and/or uses for a period of time before and/or afteradministration a compression device such as, but not limited to, acompression device which reduces the chances of deep vein thrombosis(DVT) in a subject. The compression device may be used for at least 5minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4hours, 5 hours, 6 hours, 7 hours, 8 hours, or more than 8 hours before asubject is administered the AAV particles. The compression device may beused for at least 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour,2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours,10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 1day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks ora month after the AAV particles are administered. As a non-limitingexample, the compression device is used simultaneously during theprocedure of the delivery of the AAV particles. As another non-limitingexample, the compression device is used before the administration of theAAV particles. As another non-limiting example, the compression deviceis used after administration of the AAV particles. As anothernon-limiting example, the compression device is used before, during andafter administration of the AAV particles.

Non-limiting examples, of compression devices include ActiveCare+S.F.T.intermittent compression device, ActiveCare+S.F.T pneumatic compressiondevice, DVTlite's Venowave, KCl system compression pump, AircastVenaFlow system, SCD Express Compression System or Bio CompressionSystems, Inc. pneumatic compression therapy equipment (e.g., the pumpmay be selected from Model SC-2004, Model SC-2004-FC, Model SC-3004,Model SC-3004-FC, Model SC-2008, Model SC-2008-DL, Model SC-3008-T, theBioCryo system, Model IC-BAP-DL or multi-flo DVT combo IC 1545-DL andthe garment used with the pump may be a 4 chamber, 8 chamber, BioCryo,Multi-Flo or BioArterial garment).

In one embodiment, the AAV particles may delivered to a subject using adevice to deliver the AAV particles and a head fixation assembly. Thehead fixation assembly may be, but is not limited to, any of the headfixation assemblies sold by MM interventions. As a non-limiting example,the head fixation assembly may be any of the assemblies described inU.S. Pat. Nos. 8,099,150, 8,548,569 and 9,031,636 and InternationalPatent Publication Nos. WO201108495 and WO2014014585, the contents ofeach of which are incorporated by reference in their entireties. A headfixation assembly may be used in combination with an MM compatible drillsuch as, but not limited to, the MM compatible drills described inInternational Patent Publication No. WO2013181008 and US PatentPublication No. US20130325012, the contents of which are hereinincorporated by reference in its entirety.

In one embodiment, the AAV particles may be delivered using a method,system and/or computer program for positioning apparatus to a targetpoint on a subject to deliver the AAV particles. As a non-limitingexample, the method, system and/or computer program may be the methods,systems and/or computer programs described in U.S. Pat. No. 8,340,743,the contents of which are herein incorporated by reference in itsentirety. The method may include: determining a target point in the bodyand a reference point, wherein the target point and the reference pointdefine a planned trajectory line (PTL) extending through each;determining a visualization plane, wherein the PTL intersects thevisualization plane at a sighting point; mounting the guide devicerelative to the body to move with respect to the PTL, wherein the guidedevice does not intersect the visualization plane; determining a pointof intersection (GPP) between the guide axis and the visualizationplane; and aligning the GPP with the sighting point in the visualizationplane. In one embodiment, the AAV particles may be delivered to asubject using a convention-enhanced delivery device. Non-limitingexamples of targeted delivery of drugs using convection are described inUS Patent Publication Nos. US20100217228, US20130035574 andUS20130035660 and International Patent Publication No. WO2013019830 andWO2008144585, the contents of each of which are herein incorporated byreference in their entireties.

In one embodiment, a subject may be imaged prior to, during and/or afterdelivery of the AAV particles. The imaging method may be a method knownin the art and/or described herein, such as but not limited to, magneticresonance imaging (MRI). As a non-limiting example, imaging may be usedto assess therapeutic effect. As another non-limiting example, imagingmay be used for assisted delivery of AAV particles.

In one embodiment, the AAV particles may be delivered using anMRI-guided device. Non-limiting examples of MRI-guided devices aredescribed in U.S. Pat. Nos. 9,055,884, 9,042,958, 8,886,288, 8,768,433,8,396,532, 8,369,930, 8,374,677 and 8,175,677 and US Patent

Application No. US20140024927 the contents of each of which are hereinincorporated by reference in their entireties. As a non-limitingexample, the MM-guided device may be able to provide data in real timesuch as those described in U.S. Pat. Nos. 8,886,288 and 8,768,433, thecontents of each of which is herein incorporated by reference in itsentirety. As another non-limiting example, the MRI-guided device orsystem may be used with a targeting cannula such as the systemsdescribed in U.S. Pat. Nos. 8,175,677 and 8,374,677, the contents ofeach of which are herein incorporated by reference in their entireties.As yet another non-limiting example, the MM-guided device includes atrajectory guide frame for guiding an interventional device asdescribed, for example, in U.S. Pat. No. 9,055,884 and US PatentApplication No. US20140024927, the contents of each of which are hereinincorporated by reference in their entireties.

In one embodiment the AAV particles may be delivered using anMRI-compatible tip assembly. Non-limiting examples of MRI-compatible tipassemblies are described in US Patent Publication No. US20140275980, thecontents of which is herein incorporated by reference in its entirety.

In one embodiment, the AAV particles may be delivered using a cannulawhich is MRI-compatible. Non-limiting examples of MRI-compatiblecannulas include those taught in International Patent Publication No.WO2011130107, the contents of which are herein incorporated by referencein its entirety.

In one embodiment, the AAV particles may be delivered using a catheterwhich is MRI-compatible. Non-limiting examples of MRI-compatiblecatheters include those taught in International Patent Publication No.WO2012116265, U.S. Pat. No. 8,825,133 and US Patent Publication No.US20140024909, the contents of each of which are herein incorporated byreference in their entireties.

In one embodiment, the AAV particles may be delivered using a devicewith an elongated tubular body and a diaphragm as described in US PatentPublication Nos. US20140276582 and US20140276614, the contents of eachof which are herein incorporated by reference in their entireties.

In one embodiment, the AAV particles may be delivered using an MRIcompatible localization and/or guidance system such as, but not limitedto, those described in US Patent Publication Nos. US20150223905 andUS20150230871, the contents of each of which are herein incorporated byreference in their entireties. As a non-limiting example, the MRIcompatible localization and/or guidance systems may comprise a mountadapted for fixation to a patient, a targeting cannula with a lumenconfigured to attach to the mount so as to be able to controllablytranslate in at least three dimensions, and an elongate probe configuredto snugly advance via slide and retract in the targeting cannula lumen,the elongate probe comprising at least one of a stimulation or recordingelectrode.

In one embodiment, the AAV particles may be delivered to a subject usinga trajectory frame as described in US Patent Publication Nos.US20150031982 and US20140066750 and International Patent PublicationNos. WO2015057807 and WO2014039481, the contents of each of which areherein incorporated by reference in their entireties.

In one embodiment, the AAV particles may be delivered to a subject usinga gene gun.

Definitions

At various places in the present specification, substituents ofcompounds of the present disclosure are disclosed in groups or inranges. It is specifically intended that the present disclosure includeeach and every individual subcombination of the members of such groupsand ranges.

About: As used herein, the term “about” means +/−10% of the recitedvalue.

Activity: As used herein, the term “activity” refers to the condition inwhich things are happening or being done. Compositions described hereinmay have activity and this activity may involve one or more biologicalevents.

Adeno-associated virus: The term “adeno-associated virus” or “AAV” asused herein refers to members of the dependovirus genus comprising anyparticle, sequence, gene, protein, or component derived therefrom. Theterm “AAV particle” as used herein comprises a capsid and apolynucleotide. The AAV particle may be derived from any serotype,described herein or known in the art, including combinations ofserotypes (i.e., “pseudotyped” AAV) or from various genomes (e.g.,single stranded or self-complementary). In addition, the AAV particlemay be replication defective and/or targeted.

Administered in combination: As used herein, the term “administered incombination” or “combined administration” means that two or more agents(e.g., AAV) are administered to a subject at the same time or within aninterval such that there may be an overlap of an effect of each agent onthe patient and/or the subject is at some point in time simultaneouslyexposed to both. In some embodiments, they are administered within about60, 30, 15, 10, 5, or 1 minutes of one another or within about 24 hours,12 hours, 6 hours, 3 hours of at least one dose of one or more otheragents. In some embodiments, administration occurs in overlapping dosageregimens. As used herein, the term “dosage regimen” refers to aplurality of doses spaced apart in time. Such doses may occur at regularintervals or may include one or more hiatus in administration. In someembodiments, the administrations of the agents are spaced sufficientlyclosely together such that a combinatorial (e.g., a synergistic) effectis achieved.

Amelioration: As used herein, the term “amelioration” or “ameliorating”refers to a lessening of severity of at least one indicator of acondition or disease. For example, in the context of neurodegenerationdisorder, amelioration includes the reduction of neuron loss.

Animal: As used herein, the term “animal” refers to any member of theanimal kingdom. In some embodiments, “animal” refers to humans at anystage of development. In some embodiments, “animal” refers to non-humananimals at any stage of development. In certain embodiments, thenon-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit,a monkey, a dog, a cat, a sheep, cattle, a primate, or a pig). In someembodiments, animals include, but are not limited to, mammals, birds,reptiles, amphibians, fish, and worms. In some embodiments, the animalis a transgenic animal, genetically-engineered animal, or a clone.

Antisense strand: As used herein, the term “the antisense strand” or“the first strand” or “the guide strand” of a siRNA molecule refers to astrand that is substantially complementary to a section of about 10-50nucleotides, e.g., about 15-30, 16-25, 18-23 or 19-22 nucleotides of themRNA of the gene targeted for silencing. The antisense strand or firststrand has sequence sufficiently complementary to the desired targetmRNA sequence to direct target-specific silencing, e.g., complementaritysufficient to trigger the destruction of the desired target mRNA by theRNAi machinery or process.

Approximately: As used herein, the term “approximately” or “about,” asapplied to one or more values of interest, refers to a value that issimilar to a stated reference value. In certain embodiments, the term“approximately” or “about” refers to a range of values that fall within25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%,6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than orless than) of the stated reference value unless otherwise stated orotherwise evident from the context (except where such number wouldexceed 100% of a possible value).

Associated with: As used herein, the terms “associated with,”“conjugated,” “linked,” “attached,” and “tethered,” when used withrespect to two or more moieties, means that the moieties are physicallyassociated or connected with one another, either directly or via one ormore additional moieties that serves as a linking agent, to form astructure that is sufficiently stable so that the moieties remainphysically associated under the conditions in which the structure isused, e.g., physiological conditions. An “association” need not bestrictly through direct covalent chemical bonding. It may also suggestionic or hydrogen bonding or a hybridization based connectivitysufficiently stable such that the “associated” entities remainphysically associated.

Bifunctional: As used herein, the term “bifunctional” refers to anysubstance, molecule or moiety which is capable of or maintains at leasttwo functions. The functions may affect the same outcome or a differentoutcome. The structure that produces the function may be the same ordifferent.

Biologically active: As used herein, the phrase “biologically active”refers to a characteristic of any substance (e.g., AAV) that hasactivity in a biological system and/or organism. For instance, asubstance that, when administered to an organism, has a biologicaleffect on that organism, is considered to be biologically active. Inparticular embodiments, a polynucleotide of the present disclosure maybe considered biologically active if even a portion of thepolynucleotides is biologically active or mimics an activity consideredbiologically relevant.

Biological system: As used herein, the term “biological system” refersto a group of organs, tissues, cells, intracellular components,proteins, nucleic acids, molecules (including, but not limited tobiomolecules) that function together to perform a certain biologicaltask within cellular membranes, cellular compartments, cells, tissues,organs, organ systems, multicellular organisms, or any biologicalentity. In some embodiments, biological systems are cell signalingpathways comprising intracellular and/or extracellular cell signalingbiomolecules. In some embodiments, biological systems comprise growthfactor signaling events within the extracellular/cellular matrix and/orcellular niches.

Biomolecule: As used herein, the term “biomolecule” is any naturalmolecule which is amino acid-based, nucleic acid-based,carbohydrate-based or lipid-based, and the like.

Complementary and substantially complementary: As used herein, the term“complementary” refers to the ability of polynucleotides to form basepairs with one another. Base pairs are typically formed by hydrogenbonds between nucleotide units in antiparallel polynucleotide strands.Complementary polynucleotide strands can form base pairs in theWatson-Crick manner (e.g., A to T, A to U, C to G), or in any othermanner that allows for the formation of duplexes. As persons skilled inthe art are aware, when using RNA as opposed to DNA, uracil rather thanthymine is the base that is considered to be complementary to adenosine.However, when a U is denoted in the context of the present disclosure,the ability to substitute a T is implied, unless otherwise stated.Perfect complementarity or 100% complementarity refers to the situationin which each nucleotide unit of one polynucleotide strand can form ahydrogen bond with a nucleotide unit of a second polynucleotide strand.Less than perfect complementarity refers to the situation in which some,but not all, nucleotide units of two strands can form hydrogen bondswith each other. For example, for two 20-mers, if only two base pairs oneach strand can form hydrogen bonds with each other, the polynucleotidestrands exhibit 10% complementarity. In the same example, if 18 basepairs on each strand can form hydrogen bonds with each other, thepolynucleotide strands exhibit 90% complementarity. As used herein, theterm “substantially complementary” means that the siRNA has a sequence(e.g., in the antisense strand) which is sufficient to bind the desiredtarget mRNA, and to trigger the RNA silencing of the target mRNA.

Compound: As used herein, the term “compound,” refers to a distinctchemical entity. In some embodiments, a particular compound may exist inone or more isomeric or isotopic forms (including, but not limited tostereoisomers, geometric isomers and isotopes). In some embodiments, acompound is provided or utilized in only a single such form. In someembodiments, a compound is provided or utilized as a mixture of two ormore such forms (including, but not limited to a racemic mixture ofstereoisomers). Those of skill in the art appreciate that some compoundsexist in different such forms, show different properties and/oractivities (including, but not limited to biological activities). Insuch cases it is within the ordinary skill of those in the art to selector avoid particular forms of the compound for use in accordance with thepresent disclosure. For example, compounds that contain asymmetricallysubstituted carbon atoms can be isolated in optically active or racemicforms. Methods on how to prepare optically active forms from opticallyactive starting materials are known in the art, such as by resolution ofracemic mixtures or by stereoselective synthesis. Many geometric isomersof olefins, C═N double bonds, and the like can also be present in thecompounds described herein, and all such stable isomers are contemplatedin the present disclosure. Cis and trans geometric isomers of thecompounds of the present disclosure are described and may be isolated asa mixture of isomers or as separated isomeric forms.

Compounds of the present disclosure also include tautomeric forms.Tautomeric forms result from the swapping of a single bond with anadjacent double bond and the concomitant migration of a proton.Tautomeric forms include prototropic tautomers which are isomericprotonation states having the same empirical formula and total charge.

Compounds of the present disclosure also include all of the isotopes ofthe atoms occurring in the intermediate or final compounds. “Isotopes”refers to atoms having the same atomic number but different mass numbersresulting from a different number of neutrons in the nuclei. Forexample, isotopes of hydrogen include tritium and deuterium.

The compounds and salts of the present disclosure can be prepared incombination with solvent or water molecules to form solvates andhydrates by routine methods.

Conserved: As used herein, the term “conserved” refers to nucleotides oramino acid residues of a polynucleotide sequence or polypeptidesequence, respectively, that are those that occur unaltered in the sameposition of two or more sequences being compared. Nucleotides or aminoacids that are relatively conserved are those that are conserved amongstmore related sequences than nucleotides or amino acids appearingelsewhere in the sequences.

In some embodiments, two or more sequences are said to be “completelyconserved” if they are 100% identical to one another. In someembodiments, two or more sequences are said to be “highly conserved” ifthey are at least 70% identical, at least 80% identical, at least 90%identical, or at least 95% identical to one another. In someembodiments, two or more sequences are said to be “highly conserved” ifthey are about 70% identical, about 80% identical, about 90% identical,about 95%, about 98%, or about 99% identical to one another. In someembodiments, two or more sequences are said to be “conserved” if theyare at least 30% identical, at least 40% identical, at least 50%identical, at least 60% identical, at least 70% identical, at least 80%identical, at least 90% identical, or at least 95% identical to oneanother. In some embodiments, two or more sequences are said to be“conserved” if they are about 30% identical, about 40% identical, about50% identical, about 60% identical, about 70% identical, about 80%identical, about 90% identical, about 95% identical, about 98%identical, or about 99% identical to one another. Conservation ofsequence may apply to the entire length of an oligonucleotide, apolynucleotide or polypeptide or may apply to a portion, region orfeature thereof.

In one embodiment, conserved sequences are not contiguous. Those skilledin the art are able to appreciate how to achieve alignment when gaps incontiguous alignment are present between sequences, and to aligncorresponding residues not withstanding insertions or deletions present.

In one embodiment, conserved sequences are not contiguous. Those skilledin the art are able to appreciate how to achieve alignment when gaps incontiguous alignment are present between sequences, and to aligncorresponding residues not withstanding insertions or deletions present.

Delivery: As used herein, “delivery” refers to the act or manner ofdelivering a compound such as a parvovirus, e.g. an AAV and/or AAVcompound, substance, entity, moiety, cargo or payload to a target. Suchtarget may be a cell, tissue, organ, organism, or system (whetherbiological or production).

Delivery Agent: As used herein, “delivery agent” refers to any agent orsubstance which facilitates, at least in part, the in vivo and/or invitro delivery of a polynucleotide and/or one or more substances(including, but not limited to a compounds and/or compositions of thepresent disclosure, e.g., viral particles or expression vectors) totargeted cells.

Destabilized: As used herein, the term “destable,” “destabilize,” or“destabilizing region” means a region or molecule that is less stablethan a starting, reference, wild-type or native form of the same regionor molecule.

Detectable label: As used herein, “detectable label” refers to one ormore markers, signals, or moieties which are attached, incorporated orassociated with another entity that is readily detected by methods knownin the art including radiography, fluorescence, chemiluminescence,enzymatic activity, absorbance immunological detection, and the like.Detectable labels may include radioisotopes, fluorophores, chromophores,enzymes, dyes, metal ions, ligands such as biotin, avidin, streptavidinand haptens, quantum dots, and the like. Detectable labels may belocated at any position in the entity with which they are attached,incorporated or associated. For example, when attached, incorporated inor associated with a peptide or protein, they may be within the aminoacids, the peptides, or proteins, or located at the N- or C-termini.

Dosing regimen: As used herein, a “dosing regimen” is a schedule ofadministration or physician determined regimen of treatment,prophylaxis, or palliative care.

Effective Amount: As used herein, the term “effective amount” of anagent is that amount sufficient to effect beneficial or desired results,for example, upon single or multiple dose administration to a subjectcell, in curing, alleviating, relieving or improving one or moresymptoms of a disorder, clinical results, and, as such, an “effectiveamount” depends upon the context in which it is being applied. Forexample, in the context of administering an agent that treatsParkinson's Disease, an effective amount of an agent is, for example, anamount sufficient to achieve treatment, as defined herein, ofParkinson's Disease, as compared to the response obtained withoutadministration of the agent.

Encapsulate: As used herein, the term “encapsulate” means to enclose,surround or encase.

Engineered: As used herein, embodiments are “engineered” when they aredesigned to have a feature or property, whether structural or chemical,that varies from a starting point, wild-type or native molecule. Thus,engineered agents or entities are those whose design and/or productioninclude an act of the hand of man.

Epitope: As used herein, an “epitope” refers to a surface or region on amolecule that is capable of interacting with a biomolecule. For examplea protein may contain one or more amino acids, e.g., an epitope, whichinteracts with an antibody, e.g., a biomolecule. In some embodiments,when referring to a protein or protein module, an epitope may comprise alinear stretch of amino acids or a three dimensional structure formed byfolded amino acid chains.

Expression: As used herein, “expression” of a nucleic acid sequencerefers to one or more of the following events: (1) production of an RNAtemplate from a DNA sequence (e.g., by transcription); (2) processing ofan RNA transcript (e.g., by splicing, editing, 5′ cap formation, and/or3′ end processing); (3) translation of an RNA into a polypeptide orprotein; (4) folding of a polypeptide or protein; and (5)post-translational modification of a polypeptide or protein.

Feature: As used herein, a “feature” refers to a characteristic, aproperty, or a distinctive element.

Formulation: As used herein, a “formulation” includes at least onepolynucleotide and/or compound and/or composition of the presentdisclosure (e.g., a vector, AAV particle, etc.) and a delivery agent.

Fragment: A “fragment,” as used herein, refers to a contiguous portionof a whole. For example, fragments of proteins may comprise polypeptidesobtained by digesting full-length protein isolated from cultured cells.In some embodiments, a fragment of a protein includes at least 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250 or moreamino acids. In some embodiments, fragments of an antibody includeportions of an antibody subjected to enzymatic digestion or synthesizedas such.

Functional: As used herein, a “functional” biological molecule is abiological molecule and/or entity with a structure and in a form inwhich it exhibits a property and/or activity by which it ischaracterized.

Gene expression: The term “gene expression” refers to the process bywhich a nucleic acid sequence undergoes successful transcription and inmost instances translation to produce a protein or peptide. For clarity,when reference is made to measurement of “gene expression”, this shouldbe understood to mean that measurements may be of the nucleic acidproduct of transcription, e.g., RNA or mRNA or of the amino acid productof translation, e.g., polypeptides or peptides. Methods of measuring theamount or levels of RNA, mRNA, polypeptides and peptides are well knownin the art.

Homology: As used herein, the term “homology” refers to the overallrelatedness between polymeric molecules, e.g. between nucleic acidmolecules (e.g. DNA molecules and/or RNA molecules) and/or betweenpolypeptide molecules. In some embodiments, polymeric molecules areconsidered to be “homologous” to one another if their sequences are atleast 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95%, or 99% identical or similar. The term “homologous” necessarilyrefers to a comparison between at least two sequences (polynucleotide orpolypeptide sequences). In accordance with the disclosure, twopolynucleotide sequences are considered to be homologous if thepolypeptides they encode are at least about 50%, 60%, 70%, 80%, 90%,95%, or even 99% for at least one stretch of at least about 20 aminoacids. In some embodiments, homologous polynucleotide sequences arecharacterized by the ability to encode a stretch of at least 4-5uniquely specified amino acids. For polynucleotide sequences less than60 nucleotides in length, homology is typically determined by theability to encode a stretch of at least 4-5 uniquely specified aminoacids. In accordance with the disclosure, two protein sequences areconsidered to be homologous if the proteins are at least about 50%, 60%,70%, 80%, or 90% identical for at least one stretch of at least about 20amino acids. In many embodiments, homologous protein may show a largeoverall degree of homology and a high degree of homology over at leastone short stretch of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50 or more amino acids. Inmany embodiments, homologous proteins share one or more characteristicsequence elements. As used herein, the term “characteristic sequenceelement” refers to a motif present in related proteins. In someembodiments, the presence of such motifs correlates with a particularactivity (such as biological activity).

Identity: As used herein, the term “identity” refers to the overallrelatedness between polymeric molecules, e.g., between oligonucleotideand/or polynucleotide molecules (e.g. DNA molecules and/or RNAmolecules) and/or between polypeptide molecules. Calculation of thepercent identity of two polynucleotide sequences, for example, may beperformed by aligning the two sequences for optimal comparison purposes(e.g., gaps can be introduced in one or both of a first and a secondnucleic acid sequences for optimal alignment and non-identical sequencescan be disregarded for comparison purposes). In certain embodiments, thelength of a sequence aligned for comparison purposes is at least 30%, atleast 40%, at least 50%, at least 60%, at least 70%, at least 80%, atleast 90%, at least 95%, or 100% of the length of the referencesequence. The nucleotides at corresponding nucleotide positions are thencompared. When a position in the first sequence is occupied by the samenucleotide as the corresponding position in the second sequence, thenthe molecules are identical at that position. The percent identitybetween the two sequences is a function of the number of identicalpositions shared by the sequences, taking into account the number ofgaps, and the length of each gap, which needs to be introduced foroptimal alignment of the two sequences. The comparison of sequences anddetermination of percent identity between two sequences can beaccomplished using a mathematical algorithm. For example, the percentidentity between two nucleotide sequences can be determined usingmethods such as those described in Computational Molecular Biology,Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing:Informatics and Genome Projects, Smith, D. W., ed., Academic Press, NewYork, 1993; Sequence Analysis in Molecular Biology, von Heinje, G.,Academic Press, 1987; Computer Analysis of Sequence Data, Part I,Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey,1994; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds.,M Stockton Press, New York, 1991; each of which is incorporated hereinby reference in its entirety. For example, the percent identity betweentwo nucleotide sequences can be determined, for example using thealgorithm of Meyers and Miller (CABIOS, 1989, 4:11-17), which has beenincorporated into the ALIGN program (version 2.0) using a PAM120 weightresidue table, a gap length penalty of 12 and a gap penalty of 4. Thepercent identity between two nucleotide sequences can, alternatively, bedetermined using the GAP program in the GCG software package using anNWSgapdna.CMP matrix. Methods commonly employed to determine percentidentity between sequences include, but are not limited to thosedisclosed in Carillo, H., and Lipman, D., SIAM J Applied Math., 48:1073(1988); incorporated herein by reference in its entirety. Techniques fordetermining identity are codified in publicly available computerprograms. Exemplary computer software to determine homology between twosequences include, but are not limited to, GCG program package,Devereux, J., et al., Nucleic Acids Research, 12(1), 387 (1984)),BLASTP, BLASTN, and FASTA Altschul, S. F. et al., J. Molec. Biol., 215,403 (1990)).

Inhibit expression of a gene: As used herein, the phrase “inhibitexpression of a gene” means to cause a reduction in the amount of anexpression product of the gene. The expression product may be RNAtranscribed from the gene (e.g. mRNA) or a polypeptide translated frommRNA transcribed from the gene. Typically a reduction in the level ofmRNA results in a reduction in the level of a polypeptide translatedtherefrom. The level of expression may be determined using standardtechniques for measuring mRNA or protein.

In vitro: As used herein, the term “in vitro” refers to events thatoccur in an artificial environment, e.g., in a test tube or reactionvessel, in cell culture, in a Petri dish, etc., rather than within anorganism (e.g., animal, plant, or microbe).

In vivo: As used herein, the term “in vivo” refers to events that occurwithin an organism (e.g., animal, plant, or microbe or cell or tissuethereof).

Isolated: As used herein, the term “isolated” is synonymous with“separated”, but carries with it the inference separation was carriedout by the hand of man. In one embodiment, an isolated substance orentity is one that has been separated from at least some of thecomponents with which it was previously associated (whether in nature orin an experimental setting). Isolated substances may have varying levelsof purity in reference to the substances from which they have beenassociated. Isolated substances and/or entities may be separated from atleast about 10%, about 20%, about 30%, about 40%, about 50%, about 60%,about 70%, about 80%, about 90%, or more of the other components withwhich they were initially associated. In some embodiments, isolatedagents are more than about 80%, about 85%, about 90%, about 91%, about92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%,about 99%, or more than about 99% pure. As used herein, a substance is“pure” if it is substantially free of other components.

Substantially isolated: By “substantially isolated” is meant that thecompound is substantially separated from the environment in which it wasformed or detected. Partial separation can include, for example, acomposition enriched in the compound of the present disclosure.Substantial separation can include compositions containing at leastabout 50%, at least about 60%, at least about 70%, at least about 80%,at least about 90%, at least about 95%, at least about 97%, or at leastabout 99% by weight of the compound of the present disclosure, or saltthereof. Methods for isolating compounds and their salts are routine inthe art. In some embodiments, isolation of a substance or entityincludes disruption of chemical associations and/or bonds. In someembodiments, isolation includes only the separation from components withwhich the isolated substance or entity was previously combined and doesnot include such disruption.

Modified: As used herein, the term “modified” refers to a changed stateor structure of a molecule or entity of the disclosure as compared witha parent or reference molecule or entity. Molecules may be modified inmany ways including chemically, structurally, and functionally. In someembodiments, compounds and/or compositions of the present disclosure aremodified by the introduction of non-natural amino acids, or non-naturalnucleotides.

Mutation: As used herein, the term “mutation” refers to a change and/oralteration. In some embodiments, mutations may be changes and/oralterations to proteins (including peptides and polypeptides) and/ornucleic acids (including polynucleic acids). In some embodiments,mutations comprise changes and/or alterations to a protein and/ornucleic acid sequence. Such changes and/or alterations may comprise theaddition, substitution and or deletion of one or more amino acids (inthe case of proteins and/or peptides) and/or nucleotides (in the case ofnucleic acids and or polynucleic acids). In embodiments whereinmutations comprise the addition and/or substitution of amino acidsand/or nucleotides, such additions and/or substitutions may comprise 1or more amino acid and/or nucleotide residues and may include modifiedamino acids and/or nucleotides.

Naturally occurring: As used herein, “naturally occurring” meansexisting in nature without artificial aid or involvement of the hand ofman

Non-human vertebrate: As used herein, a “non-human vertebrate” includesall vertebrates except Homo sapiens, including wild and domesticatedspecies. Examples of non-human vertebrates include, but are not limitedto, mammals, such as alpaca, banteng, bison, camel, cat, cattle, deer,dog, donkey, gayal, goat, guinea pig, horse, llama, mule, pig, rabbit,reindeer, sheep water buffalo, and yak.

Nucleic acid: As used herein, the term “nucleic acid”, “polynucleotide”and ‘oligonucleotide” refer to any nucleic acid polymers composed ofeither polydeoxyribonucleotides (containing 2-deoxy-D-ribose), orpolyribonucleotides (containing D-ribose), or any other type ofpolynucleotide which is an N glycoside of a purine or pyrimidine base,or modified purine or pyrimidine bases. There is no intended distinctionin length between the term “nucleic acid”, “polynucleotide” and“oligonucleotide”, and these terms will be used interchangeably. Theseterms refer only to the primary structure of the molecule. Thus, theseterms include double- and single-stranded DNA, as well as double- andsingle stranded RNA.

Off-target: As used herein, “off target” refers to any unintended effecton any one or more target, gene and/or cellular transcript.

Open reading frame: As used herein, “open reading frame” or “ORF” refersto a sequence which does not contain a stop codon in a given readingframe.

Operably linked: As used herein, the phrase “operably linked” refers toa functional connection between two or more molecules, constructs,transcripts, entities, moieties or the like.

Particle: As used herein, a “particle” is a virus comprised of at leasttwo components, a protein capsid and a polynucleotide sequence enclosedwithin the capsid.

Patient: As used herein, “patient” refers to a subject who may seek orbe in need of treatment, requires treatment, is receiving treatment,will receive treatment, or a subject who is under care by a trainedprofessional for a particular disease or condition, such as for exampleParkinson's Disease.

Payload: As used herein, “payload” refers to one or more polynucleotidesor polynucleotide regions encoded by or within a viral genome or anexpression product of such polynucleotide or polynucleotide region,e.g., a transgene, a polynucleotide encoding a polypeptide ormulti-polypeptide or a modulatory nucleic acid or regulatory nucleicacid.

Payload construct: As used herein, “payload construct” is one or morepolynucleotide regions encoding or comprising a payload that is flankedon one or both sides by an inverted terminal repeat (ITR) sequence. Thepayload construct is a template that is replicated in a viral productioncell to produce a viral genome.

Payload construct vector: As used herein, “payload construct vector” isa vector encoding or comprising a payload construct, and regulatoryregions for replication and expression in bacterial cells.

Payload construct expression vector: As used herein, a “payloadconstruct expression vector” is a vector encoding or comprising apayload construct and which further comprises one or more polynucleotideregions encoding or comprising components for viral expression in aviral replication cell.

Peptide: As used herein, “peptide” is less than or equal to 50 aminoacids long, e.g., about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 aminoacids long.

Pharmaceutically acceptable: The phrase “pharmaceutically acceptable” isemployed herein to refer to those compounds, materials, compositions,and/or dosage forms which are, within the scope of sound medicaljudgment, suitable for use in contact with the tissues of human beingsand animals without excessive toxicity, irritation, allergic response,or other problem or complication, commensurate with a reasonablebenefit/risk ratio.

Pharmaceutically acceptable excipients: The phrase “pharmaceuticallyacceptable excipient,” as used herein, refers to any ingredient otherthan the compounds and/or active agents (e.g. as described herein)present in pharmaceutical compositions and having the properties ofbeing substantially nontoxic and non-inflammatory in a subject such as apatient. In some embodiments, pharmaceutically acceptable excipients arevehicles capable of suspending and/or dissolving active agents.Excipients may include, for example: antiadherents, antioxidants,binders, coatings, compression aids, disintegrants, dyes (colors),emollients, emulsifiers, fillers (diluents), film formers or coatings,flavors, fragrances, glidants (flow enhancers), lubricants,preservatives, printing inks, sorbents, suspension or dispersing agents,sweeteners, and waters of hydration. Exemplary excipients include, butare not limited to: butylated hydroxytoluene (BHT), calcium carbonate,calcium phosphate (dibasic), calcium stearate, croscarmellose,crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine,ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol,methionine, methylcellulose, methyl paraben, microcrystalline cellulose,polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinizedstarch, propyl paraben, retinyl palmitate, shellac, silicon dioxide,sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate,sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide,vitamin A, vitamin E, vitamin C, and xylitol.

Pharmaceutically acceptable salts: Pharmaceutically acceptable salts ofthe compounds described herein. As used herein, “pharmaceuticallyacceptable salts” refers to derivatives or forms of the disclosedcompounds wherein the parent compound is modified by converting anexisting acid or base moiety to its salt form (e.g., as generated byreacting the free base group with a suitable organic acid). Examples ofpharmaceutically acceptable salts include, but are not limited to,mineral or organic acid salts of basic residues such as amines; alkalior organic salts of acidic residues such as carboxylic acids; and thelike. Representative acid addition salts include acetate, acetic acid,adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzenesulfonic acid, benzoate, bisulfate, borate, butyrate, camphorate,camphorsulfonate, citrate, cyclopentanepropionate, digluconate,dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate,glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide,hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like, aswell as nontoxic ammonium, quaternary ammonium, and amine cations,including, but not limited to ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, ethylamine, and the like. The pharmaceutically acceptablesalts of the present disclosure include the conventional non-toxic saltsof the parent compound formed, for example, from non-toxic inorganic ororganic acids. In some embodiments a pharmaceutically acceptable salt ofthe present disclosure can be synthesized salt prepared from the parentcompound which contains a basic or acidic moiety by conventionalchemical methods. Generally, such salts can be prepared by reacting thefree acid or base forms of these compounds with a stoichiometric amountof the appropriate base or acid in water or in an organic solvent, or ina mixture of the two; generally, nonaqueous media like ether, ethylacetate, ethanol, isopropanol, or acetonitrile are preferred. Lists ofsuitable salts are found in Remington's Pharmaceutical Sciences, 17thed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, PharmaceuticalSalts: Properties, Selection, and Use, P.H. Stahl and C.G. Wermuth(eds.), Wiley-VCH, 2008, and Berge et al., Journal of PharmaceuticalScience, 66, 1-19 (1977), each of which is incorporated herein byreference in its entirety.

Pharmaceutically acceptable solvate: The term “pharmaceuticallyacceptable solvate,” as used herein, refers to a crystalline form of acompound of the disclosure wherein molecules of a suitable solvent areincorporated in the crystal lattice. A suitable solvent isphysiologically tolerable at the dosage administered. For example,solvates may be prepared by crystallization, recrystallization, orprecipitation from a solution that includes organic solvents, water, ora mixture thereof. Examples of suitable solvents are ethanol, water (forexample, mono-, di-, and tri-hydrates), N-methylpyrrolidinone (NMP),dimethyl sulfoxide (DMSO), N,N′-dimethylformamide (DMF),N,N′-dimethylacetamide (DMAC), 1,3-dimethyl-2-imidazolidinone (DMEU),1,3-dimethyl-3,4,5,6-tetrahydro-2-(1H)-pyrimidinone (DMPU), acetonitrile(ACN), propylene glycol, ethyl acetate, benzyl alcohol, 2-pyrrolidone,benzyl benzoate, and the like. When water is the solvent, the solvate isreferred to as a “hydrate.” In some embodiments, the solventincorporated into a solvate is of a type or at a level that isphysiologically tolerable to an organism to which the solvate isadministered (e.g., in a unit dosage form of a pharmaceuticalcomposition).

Pharmacokinetic: As used herein, “pharmacokinetic” refers to any one ormore properties of a molecule or compound as it relates to thedetermination of the fate of substances administered to a livingorganism. Pharmacokinetics is divided into several areas including theextent and rate of absorption, distribution, metabolism and excretion.This is commonly referred to as ADME where: (A) Absorption is theprocess of a substance entering the blood circulation; (D) Distributionis the dispersion or dissemination of substances throughout the fluidsand tissues of the body; (M) Metabolism (or Biotransformation) is theirreversible transformation of parent compounds into daughtermetabolites; and (E) Excretion (or Elimination) refers to theelimination of the substances from the body. In rare cases, some drugsirreversibly accumulate in body tissue.

Physicochemical: As used herein, “physicochemical” means of or relatingto a physical and/or chemical property.

Preventing: As used herein, the term “preventing” refers to partially orcompletely delaying onset of an infection, disease, disorder and/orcondition; partially or completely delaying onset of one or moresymptoms, features, or clinical manifestations of a particularinfection, disease, disorder, and/or condition; partially or completelydelaying onset of one or more symptoms, features, or manifestations of aparticular infection, disease, disorder, and/or condition; partially orcompletely delaying progression from an infection, a particular disease,disorder and/or condition; and/or decreasing the risk of developingpathology associated with the infection, the disease, disorder, and/orcondition, such as for example Parkinson's Disease.

Prodrug: The present disclosure also includes prodrugs of the compoundsdescribed herein. As used herein, “prodrugs” refer to any substance,molecule or entity which is in a form predicate for that substance,molecule or entity to act as a therapeutic upon chemical or physicalalteration. Prodrugs may by covalently bonded or sequestered in some wayand which release or are converted into the active drug moiety prior to,upon or after administered to a mammalian subject. Preparation and useof prodrugs is discussed in T. Higuchi and V. Stella, “Pro-drugs asNovel Delivery Systems,” Vol. 14 of the A.C.S. Symposium Series, and inBioreversible Carriers in Drug Design, ed. Edward B. Roche, AmericanPharmaceutical Association and Pergamon Press, 1987, both of which arehereby incorporated by reference in their entirety.

Proliferate: As used herein, the term “proliferate” means to grow,expand, replicate or increase or cause to grow, expand, replicate orincrease. “Proliferative” means having the ability to proliferate.“Anti-proliferative” means having properties counter to or in oppositionto proliferative properties.

Prophylactic: As used herein, “prophylactic” refers to a therapeutic orcourse of action used to prevent the spread of disease.

Prophylaxis: As used herein, a “prophylaxis” refers to a measure takento maintain health and prevent the spread of disease.

Protein of interest: As used herein, the terms “proteins of interest” or“desired proteins” include those provided herein and fragments, mutants,variants, and alterations thereof.

Purified: As used herein, “purify,” “purified,” “purification” means tomake substantially pure or clear from unwanted components, materialdefilement, admixture or imperfection. “Purified” refers to the state ofbeing pure. “Purification” refers to the process of making pure.

Region: As used herein, the term “region” refers to a zone or generalarea. In some embodiments, when referring to a protein or proteinmodule, a region may comprise a linear sequence of amino acids along theprotein or protein module or may comprise a three dimensional area, anepitope and/or a cluster of epitopes. In some embodiments, regionscomprise terminal regions. As used herein, the term “terminal region”refers to regions located at the ends or termini of a given agent. Whenreferring to proteins, terminal regions may comprise N- and/orC-termini. N-termini refer to the end of a protein comprising an aminoacid with a free amino group. C-termini refer to the end of a proteincomprising an amino acid with a free carboxyl group. N- and/orC-terminal regions may therefore comprise the N- and/or C-termini aswell as surrounding amino acids. In some embodiments, N- and/orC-terminal regions comprise from about 3 amino acid to about 30 aminoacids, from about 5 amino acids to about 40 amino acids, from about 10amino acids to about 50 amino acids, from about 20 amino acids to about100 amino acids and/or at least 100 amino acids. In some embodiments,N-terminal regions may comprise any length of amino acids that includesthe N-terminus, but does not include the C-terminus. In someembodiments, C-terminal regions may comprise any length of amino acids,which include the C-terminus, but do not comprise the N-terminus.

In some embodiments, when referring to a polynucleotide, a region maycomprise a linear sequence of nucleic acids along the polynucleotide ormay comprise a three dimensional area, secondary structure, or tertiarystructure. In some embodiments, regions comprise terminal regions. Asused herein, the term “terminal region” refers to regions located at theends or termini of a given agent. When referring to polynucleotides,terminal regions may comprise 5′ and 3′ termini. 5′ termini refer to theend of a polynucleotide comprising a nucleic acid with a free phosphategroup. 3′ termini refer to the end of a polynucleotide comprising anucleic acid with a free hydroxyl group. 5′ and 3′ regions may thereforecomprise the 5′ and 3′ termini as well as surrounding nucleic acids. Insome embodiments, 5′ and 3′ terminal regions comprise from about 9nucleic acids to about 90 nucleic acids, from about 15 nucleic acids toabout 120 nucleic acids, from about 30 nucleic acids to about 150nucleic acids, from about 60 nucleic acids to about 300 nucleic acidsand/or at least 300 nucleic acids. In some embodiments, 5′ regions maycomprise any length of nucleic acids that includes the 5′ terminus, butdoes not include the 3′ terminus. In some embodiments, 3′ regions maycomprise any length of nucleic acids, which include the 3′ terminus, butdoes not comprise the 5′ terminus.

RNA or RNA molecule: As used herein, the term “RNA” or “RNA molecule” or“ribonucleic acid molecule” refers to a polymer of ribonucleotides; theterm “DNA” or “DNA molecule” or “deoxyribonucleic acid molecule” refersto a polymer of deoxyribonucleotides. DNA and RNA can be synthesizednaturally, e.g., by DNA replication and transcription of DNA,respectively; or be chemically synthesized. DNA and RNA can besingle-stranded (i.e., ssRNA or ssDNA, respectively) or multi-stranded(e.g., double stranded, i.e., dsRNA and dsDNA, respectively). The term“mRNA” or “messenger RNA”, as used herein, refers to a single strandedRNA that encodes the amino acid sequence of one or more polypeptidechains.

RNA interference: As used herein, the term “RNA interference” or “RNAi”refers to a sequence specific regulatory mechanism mediated by RNAmolecules which results in the inhibition or interference or “silencing”of the expression of a corresponding protein-coding gene.

Sample: As used herein, the term “sample” refers to an aliquot, subsetor portion taken from a source and/or provided for analysis orprocessing. In some embodiments, a sample is from a biological sourcesuch as a tissue, cell or component part (e.g. a body fluid, includingbut not limited to blood, mucus, lymphatic fluid, synovial fluid,cerebrospinal fluid, saliva, amniotic fluid, amniotic cord blood, urine,vaginal fluid and semen). In some embodiments, a sample may be orcomprise a homogenate, lysate or extract prepared from a whole organismor a subset of its tissues, cells or component parts, or a fraction orportion thereof, including but not limited to, for example, plasma,serum, spinal fluid, lymph fluid, the external sections of the skin,respiratory, intestinal, and genitourinary tracts, tears, saliva, milk,blood cells, tumors, or organs. In some embodiments, a sample is orcomprises a medium, such as a nutrient broth or gel, which may containcellular components, such as proteins or nucleic acid molecule. In someembodiments, a “primary” sample is an aliquot of the source. In someembodiments, a primary sample is subjected to one or more processing(e.g., separation, purification, etc.) steps to prepare a sample foranalysis or other use.

Self-complementary viral particle: As used herein, a “self-complementaryviral particle” is a particle comprised of at least two components, aprotein capsid and a polynucleotide sequence encoding aself-complementary genome enclosed within the capsid.

Sense strand: As used herein, the term “the sense strand” or “the secondstrand” or “the passenger strand” of a siRNA molecule refers to a strandthat is complementary to the antisense strand or first strand. Theantisense and sense strands of a siRNA molecule are hybridized to form aduplex structure. As used herein, a “siRNA duplex” includes a siRNAstrand having sufficient complementarity to a section of about 10-50nucleotides of the mRNA of the gene targeted for silencing and a siRNAstrand having sufficient complementarity to form a duplex with the siRNAstrand.

Signal Sequences: As used herein, the phrase “signal sequences” refersto a sequence which can direct the transport or localization.

Single unit dose: As used herein, a “single unit dose” is a dose of anytherapeutic administered in one dose/at one time/single route/singlepoint of contact, i.e., single administration event. In someembodiments, a single unit dose is provided as a discrete dosage form(e.g., a tablet, capsule, patch, loaded syringe, vial, etc.).

Similarity: As used herein, the term “similarity” refers to the overallrelatedness between polymeric molecules, e.g. between polynucleotidemolecules (e.g. DNA molecules and/or RNA molecules) and/or betweenpolypeptide molecules. Calculation of percent similarity of polymericmolecules to one another can be performed in the same manner as acalculation of percent identity, except that calculation of percentsimilarity takes into account conservative substitutions as isunderstood in the art.

Small/short interfering RNA: As used herein, the term “small/shortinterfering RNA” or “siRNA” refers to an RNA molecule (or RNA analog)comprising between about 5-60 nucleotides (or nucleotide analogs) whichis capable of directing or mediating RNAi. Preferably, a siRNA moleculecomprises between about 15-30 nucleotides or nucleotide analogs, morepreferably between about 16-25 nucleotides (or nucleotide analogs), evenmore preferably between about 18-23 nucleotides (or nucleotide analogs),and even more preferably between about 19-22 nucleotides (or nucleotideanalogs) (e.g., 19, 20, 21 or 22 nucleotides or nucleotide analogs). Theterm “short” siRNA refers to a siRNA comprising 5-23 nucleotides,preferably 21 nucleotides (or nucleotide analogs), for example, 19, 20,21 or 22 nucleotides. The term “long” siRNA refers to a siRNA comprising24-60 nucleotides, preferably about 24-25 nucleotides, for example, 23,24, 25 or 26 nucleotides. Short siRNAs may, in some instances, includefewer than 19 nucleotides, e.g., 16, 17 or 18 nucleotides, or as few as5 nucleotides, provided that the shorter siRNA retains the ability tomediate RNAi. Likewise, long siRNAs may, in some instances, include morethan 26 nucleotides, e.g., 27, 28, 29, 30, 35, 40, 45, 50, 55, or even60 nucleotides, provided that the longer siRNA retains the ability tomediate RNAi or translational repression absent further processing,e.g., enzymatic processing, to a short siRNA. siRNAs can be singlestranded RNA molecules (ss-siRNAs) or double stranded RNA molecules(ds-siRNAs) comprising a sense strand and an antisense strand whichhybridized to form a duplex structure called siRNA duplex.

Split dose: As used herein, a “split dose” is the division of singleunit dose or total daily dose into two or more doses.

Stable: As used herein “stable” refers to a compound or entity that issufficiently robust to survive isolation to a useful degree of purityfrom a reaction mixture, and preferably capable of formulation into anefficacious therapeutic agent.

Stabilized: As used herein, the term “stabilize”, “stabilized,”“stabilized region” means to make or become stable. In some embodiments,stability is measured relative to an absolute value. In someembodiments, stability is measured relative to a reference compound orentity.

Subject: As used herein, the term “subject” or “patient” refers to anyorganism to which a composition in accordance with the disclosure may beadministered, e.g., for experimental, diagnostic, prophylactic, and/ortherapeutic purposes. Typical subjects include animals (e.g., mammalssuch as mice, rats, rabbits, non-human primates, and humans) and/orplants. In some embodiments, the subject may be an infant, neonate, or achild under the age of 12 years old. In some embodiments, the subjectmay be in utero.

Substantially: As used herein, the term “substantially” refers to thequalitative condition of exhibiting total or near-total extent or degreeof a characteristic or property of interest. One of ordinary skill inthe biological arts will understand that biological and chemicalphenomena rarely, if ever, go to completion and/or proceed tocompleteness or achieve or avoid an absolute result. The term“substantially” is therefore used herein to capture the potential lackof completeness inherent in many biological and chemical phenomena.

Substantially equal: As used herein as it relates to time differencesbetween doses, the term means plus/minus 2%.

Substantially simultaneously: As used herein and as it relates toplurality of doses, the term typically means within about 2 seconds.

Suffering from: An individual who is “suffering from” a disease,disorder, and/or condition has been diagnosed with or displays one ormore symptoms of a disease, disorder, and/or condition such as forexample Parkinson's Disease.

Susceptible to: An individual who is “susceptible to” a disease,disorder, and/or condition has not been diagnosed with and/or may notexhibit symptoms of the disease, disorder, and/or condition but harborsa propensity to develop a disease or its symptoms. In some embodiments,an individual who is susceptible to a disease, disorder, and/orcondition (for example, cancer) may be characterized by one or more ofthe following: (1) a genetic mutation associated with development of thedisease, disorder, and/or condition; (2) a genetic polymorphismassociated with development of the disease, disorder, and/or condition;(3) increased and/or decreased expression and/or activity of a proteinand/or nucleic acid associated with the disease, disorder, and/orcondition; (4) habits and/or lifestyles associated with development ofthe disease, disorder, and/or condition; (5) a family history of thedisease, disorder, and/or condition; and (6) exposure to and/orinfection with a microbe associated with development of the disease,disorder, and/or condition. In some embodiments, an individual who issusceptible to a disease, disorder, and/or condition will develop thedisease, disorder, and/or condition. In some embodiments, an individualwho is susceptible to a disease, disorder, and/or condition will notdevelop the disease, disorder, and/or condition.

Sustained release: As used herein, the term “sustained release” refersto a pharmaceutical composition or compound release profile thatconforms to a release rate over a specific period of time.

Synthetic: The term “synthetic” means produced, prepared, and/ormanufactured by the hand of man. Synthesis of polynucleotides orpolypeptides or other molecules of the present disclosure may bechemical or enzymatic.

Targeting: As used herein, “targeting” means the process of design andselection of nucleic acid sequence that will hybridize to a targetnucleic acid and induce a desired effect.

Targeted Cells: As used herein, “targeted cells” refers to any one ormore cells of interest. The cells may be found in vitro, in vivo, insitu or in the tissue or organ of an organism. The organism may be ananimal, preferably a mammal, more preferably a human and most preferablya patient.

Therapeutic Agent: The term “therapeutic agent” refers to any agentthat, when administered to a subject, has a therapeutic, diagnostic,and/or prophylactic effect and/or elicits a desired biological and/orpharmacological effect.

Therapeutically effective amount: As used herein, the term“therapeutically effective amount” means an amount of an agent to bedelivered (e.g., nucleic acid, drug, therapeutic agent, diagnosticagent, prophylactic agent, etc.) that is sufficient, when administeredto a subject suffering from or susceptible to an infection, disease,disorder, and/or condition, to treat, improve symptoms of, diagnose,prevent, and/or delay the onset of the infection, disease, disorder,and/or condition such as for example Parkinson's Disease. In someembodiments, a therapeutically effective amount is provided in a singledose. In some embodiments, a therapeutically effective amount isadministered in a dosage regimen comprising a plurality of doses. Thoseskilled in the art will appreciate that in some embodiments, a unitdosage form may be considered to comprise a therapeutically effectiveamount of a particular agent or entity if it comprises an amount that iseffective when administered as part of such a dosage regimen.

Therapeutically effective outcome: As used herein, the term“therapeutically effective outcome” means an outcome that is sufficientin a subject suffering from or susceptible to an infection, disease,disorder, and/or condition, to treat, improve symptoms of, diagnose,prevent, and/or delay the onset of the infection, disease, disorder,and/or condition.

Total daily dose: As used herein, a “total daily dose” is an amountgiven or prescribed in 24 hour period. It may be administered as asingle unit dose.

Transfection: As used herein, the term “transfection” refers to methodsto introduce exogenous nucleic acids into a cell. Methods oftransfection include, but are not limited to, chemical methods, physicaltreatments and cationic lipids or mixtures.

Treating: As used herein, the term “treating” refers to partially orcompletely alleviating, ameliorating, improving, relieving, delayingonset of, inhibiting progression of, reducing severity of, and/orreducing incidence of one or more symptoms or features of a particularinfection, disease, disorder, and/or condition. For example, “treating”cancer may refer to inhibiting survival, growth, and/or spread of atumor. Treatment may be administered to a subject who does not exhibitsigns of a disease, disorder, and/or condition and/or to a subject whoexhibits only early signs of a disease, disorder, and/or condition forthe purpose of decreasing the risk of developing pathology associatedwith the disease, disorder, and/or condition such as for exampleParkinson's Disease.

Unmodified: As used herein, “unmodified” refers to any substance,compound or molecule prior to being changed in any way. Unmodified may,but does not always, refer to the wild type or native form of abiomolecule or entity. Molecules or entities may undergo a series ofmodifications whereby each modified substance, compound, molecule orentity may serve as the “unmodified” starting molecule for a subsequentmodification.

Vector: As used herein, a “vector” is any molecule or moiety whichtransports, transduces or otherwise acts as a carrier of a heterologousmolecule. Vectors of the present disclosure may be producedrecombinantly and may be based on and/or may comprise adeno-associatedvirus (AAV) parent or reference sequence. Such parent or reference AAVsequences may serve as an original, second, third or subsequent sequencefor engineering vectors. In non-limiting examples, such parent orreference AAV sequences may comprise any one or more of the followingsequences: a polynucleotide sequence encoding a polypeptide ormulti-polypeptide, which sequence may be wild-type or modified fromwild-type and which sequence may encode full-length or partial sequenceof a protein, protein domain, or one or more subunits of a protein; apolynucleotide comprising a modulatory or regulatory nucleic acid whichsequence may be wild-type or modified from wild-type; and a transgenethat may or may not be modified from wild-type sequence. These AAVsequences may serve as either the “donor” sequence of one or more codons(at the nucleic acid level) or amino acids (at the polypeptide level) or“acceptor” sequences of one or more codons (at the nucleic acid level)or amino acids (at the polypeptide level).

Viral construct vector: As used herein, a “viral construct vector” is avector which comprises one or more polynucleotide regions encoding orcomprising Rep and or Cap protein.

Viral construct expression vector: As used herein, a “viral constructexpression vector” is a vector which comprises one or morepolynucleotide regions encoding or comprising Rep and or Cap thatfurther comprises one or more polynucleotide regions encoding orcomprising components for viral expression in a viral replication cell.

Viral genome: As used herein, a “viral genome” is a polynucleotideencoding at least one inverted terminal repeat (ITR), at least oneregulatory sequence, and at least one payload. The viral genome isderived by replication of a payload construct from the payload constructexpression vector. A viral genome encodes at least one copy of thepayload construct.

EQUIVALENTS AND SCOPE

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments in accordance with the disclosure described herein. Thescope of the present disclosure is not intended to be limited to theabove Description, but rather is as set forth in the appended claims.

In the claims, articles such as “a,” “an,” and “the” may mean one ormore than one unless indicated to the contrary or otherwise evident fromthe context. Claims or descriptions that include “or” between one ormore members of a group are considered satisfied if one, more than one,or all of the group members are present in, employed in, or otherwiserelevant to a given product or process unless indicated to the contraryor otherwise evident from the context. The disclosure includesembodiments in which exactly one member of the group is present in,employed in, or otherwise relevant to a given product or process. Thedisclosure includes embodiments in which more than one, or all of thegroup members are present in, employed in, or otherwise relevant to agiven product or process.

It is also noted that the term “comprising” is intended to be open andpermits but does not require the inclusion of additional elements orsteps. When the term “comprising” is used herein, the term “consistingof” is thus also encompassed and disclosed.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure belongs. Methods and materials aredescribed herein for use in the present disclosure; other, suitablemethods and materials known in the art can also be used.

Where ranges are given, endpoints are included. Furthermore, it is to beunderstood that unless otherwise indicated or otherwise evident from thecontext and understanding of one of ordinary skill in the art, valuesthat are expressed as ranges can assume any specific value or subrangewithin the stated ranges in different embodiments of the disclosure, tothe tenth of the unit of the lower limit of the range, unless thecontext clearly dictates otherwise.

In addition, it is to be understood that any particular embodiment ofthe present disclosure that falls within the prior art may be explicitlyexcluded from any one or more of the claims. Since such embodiments aredeemed to be known to one of ordinary skill in the art, they may beexcluded even if the exclusion is not set forth explicitly herein. Anyparticular embodiment of the compositions of the disclosure (e.g., anynucleic acid or protein encoded thereby; any method of production; anymethod of use; etc.) can be excluded from any one or more claims, forany reason, whether or not related to the existence of prior art.

All cited sources, for example, references, publications, databases,database entries, and art cited herein, are incorporated into thisapplication by reference, even if not expressly stated in the citation.In case of conflicting statements of a cited source and the instantapplication, the statement in the instant application shall control.

Section and table headings are not intended to be limiting.

EXAMPLES Example 1. Design of Payloads: AADC Polynucleotides

AADC polynucleotides are designed to comprise at a minimum a nucleicacid sequence encoding an AADC protein.

Once designed, the sequence is engineered or synthesized or inserted ina plasmid or vector and administered to a cell or organism. Suitableplasmids or vectors are any which transduce or transfect the targetcell.

Adeno-associated viral (AAV) particles may be used.

Administration results in the processing of the AADC polynucleotide togenerate the AADC protein which alters the etiology of the disease, inthis case Parkinson's Disease. In one non-limiting example, plasmidscontaining an AADC polynucleotide of the disclosure have a CMV promoterand encode AADC. In some embodiments the open reading frame of the AADCprotein mRNA is codon optimized.

AADC polynucleotides, listed ITR to ITR, suitable for use in a AAVparticles include those in Table 2.

TABLE 2 ITR to ITR AADC polynucleotides Construct SEQ ID NO AADCPolynucleotide 979

The start and stop positions of various regions of the AADCpolynucleotides are given are relative to the ITR to ITR AADCpolynucleotides described in Table 2. In Table 3, ITR is invertedterminal repeat, MCS is multiple cloning site, CMV is cytomegalovirus,Ie1 is immediate-early 1, hBglobin is human beta-globin, AADC is regionencoding the AADC polypeptide, and poly(A) is the polyadenylationsignal.

TABLE 3 Component regions of AADC polynucleotides AADC Polynucleotide(SEQ ID NO: 979) Length SEQ ID NO Region Start Stop of Region of region5′ ITR 1 141 141 980 MCS 189 206 18 981 CMV enhancer 213 515 303 982 CMVpromoter 516 719 204 983 Ie1 exon 1 734 867 134 984 Ie1 intron partial868 899 32 985 hBglobin intron 2 900 1246 347 986 hBglobin exon 3 12471299 53 987 AADC 1338 2777 1440 988 MCS 2820 2837 18 989 Poly(A) 28383314 477 990 3′ ITR 3386 3526 141 991

Example 2. Design of AADC Polynucleotides to Treat Parkinson's Disease

AADC polynucleotides are designed to comprise at a minimum a nucleicacid sequence encoding an AADC protein.

Once designed, the sequence is engineered or synthesized or inserted ina plasmid or vector and administered to a cell or organism. Suitableplasmids or vectors are any which transduce or transfect the targetcell.

Adeno-associated viral (AAV) particles may be used.

Administration results in the processing of the AADC polynucleotide togenerate the AADC protein which alters the etiology of the disease, inthis case Parkinson's Disease.

Example 3. Administration of AAV Particles

AAV particles are infused into the substantia nigra, and in particular,the substantia nigra pars compacta (SNpc) and ventral tegmental area(VTA) of patients having Parkinson's Disease and identified as qualifiedfor treatment according to methods known in the art.

One method of administration contemplated for use in the methodsdescribed herein is real-time convection-enhanced delivery (RCD) of AAVparticle compositions by co-infusion of gadoteridol (a magneticresonance (MR) contrast agent) and T1 or T2 magnetic resonance imaging(MRI), which can predict areas of subsequent AADC gene expression. Asdescribed in Richardson, et al., 2011, the accuracy of cannula placementand initial infusate distribution may be safely determined by salineinfusion without significantly altering the subsequent distribution ofthe tracer agent (Richardson, et al., 2011, Neurosurgery,69(1):154-163). T2 RCD provides detection of intraparenchymalconvection-enhanced delivery in the uninjured brain and may predictsubsequent distribution of a transgene after AAV particle infusion.Subjects undergo saline infusion/T2 acquisition, immediately followed bygadoteridol infusion/T1 acquisition in the putamen and brainstem.Distribution volumes and spatial patterns are analyzed. Gadoteridol andAAV-encoded AADC are co-infused under alternating T2/T1 acquisition inthe thalamus, and hyperintense areas are compared with areas ofsubsequent transgene expression. Ratios of distribution volume toinfusion volume are expected to be similar between saline andgadoteridol RCD. Spatial overlap should correlate well between T2 and T1images. The second infusate will follow a spatiotemporal pattern similarto that of the first, filling the target area before developingextra-target distribution. Areas of AADC expression should correlatewell with areas of both T1 and T2 hyperintensity observed during RCD(Richardson, et al., 2011, Neurosurgery, 69(1):154-163).

Convection-enhanced delivery (CED) of macromolecules directly into thebrain parenchyma has been known for over two decades. CED is a term thatdenotes the use of a pressure gradient to generate bulk flow within thebrain parenchyma, i.e. convection of macromolecules within theinterstitial fluid driven by infusing a solution through a cannulaplaced directly in the targeted structure. This method allowstherapeutic agents to be homogenously distributed through large volumesof brain tissue by bypassing the blood brain barrier and surpassingsimple diffusion (Richardson, et al., 2011, Stereotact. Funct.Neurosurg. 89:141-151).

Salegio, et al. recently demonstrated the distribution of nanoparticlesof different sizes, including micelles (˜15 nm in size), AAV (˜20-25 nm)and liposomes (˜65 nm), within the CNS of rodents and NHPs (Salegio etal., 2014, Frontiers in Neuroanatomy, vol. 8, article 9: pp. 1-8).Simple injections cannot engage the perivascular system, and specializedinfusion cannulae are required, enabling constant pressures to beexerted at the tip of the cannula such that the interstitial hydrostaticpressure is exceeded and infusate can flow out into the tissue. Simpleneedles generate significant reflux; thus, reflux-resistant cannulashave been developed to counter this tendency. The advent of platformsfor MM-guided convection-enhanced infusions further refinedunderstanding of the mechanics of perivascular flow, and it wasdemonstrated that perivascular distribution of liposomes was linear withrespect to time, the slope of the curve was increased in myelinatedregions, and cessation of infusion prevented further expansion in thevolume of distribution. (Richardson, et al., 2011, Stereotact. Funct.Neurosurg. 89:141-151; Salegio et al., 2014, Frontiers in Neuroanatomy,vol. 8, article 9: pp. 1-8).

Intraparenchymal rAAV injections are known to result in robust butrelatively local transduction. Such local delivery methods areadvantageous when attempting gene therapy for neurological disordersthat result from neuropathology that is localized to a specificanatomical region or anatomical circuitry such as in the case ofParkinson's disease. However, in treatments requiring more widespreadCNS transduction, intraparenchymal injections are impractical. Treatmentof neurological disorders attributable to inborn errors of metabolismand/or single-gene defects, or those that affect motor neurons of thespinal cord can require transduction of large proportions of the brainor spinal cord, respectively. Development of less invasive trans-BBBdelivery methods for vectors is an extremely important endeavor.Numerous attempts to use molecules that are known to interact withvarious active transport mechanisms (probably receptor-mediated) toconvey proteins across the BBB have been reported with varying results.Given the large number of AAV serotypes available, one or more serotypesmay bind a cell-entry receptor capable of transporting the AAV capsidacross the BBB (Manfredsson, et al., 2009, “AAV9: a potentialblood-brain barrier buster.” Molecular Therapy 17(3):403-405).

Vector and Stereotaxic Infusion

A stereotactic approach may be used to surgically deliver the AADCpolynucleotides. Although individuals with AADC deficiency lackepinephrine and norepinephrine, these patients should maintain stableblood pressure and heart rates during the surgery. There should be nonotable intracerebral hemorrhages in the postoperative computedtomography (CT) or MRI scans. The needle tracts, as shown on the MMscans, should show accurate injection into the substantia nigra parscompacta (SNpc) and ventral tegmental area (VTA). The patients will bedischarged from the hospital about one week after the surgery (Hwu, W.L., et al., 2012. Gene therapy for aromatic L-amino acid decarboxylasedeficiency. Sci. Transl. Med. Vol. 4, 134ra61).

Subjects of treatment receive the AAV-vector composition vector, safelydelivered to substantia nigra pars compacta (SNpc) and ventral tegmentalarea (VTA) via bilateral infusions, or alternatively, intrastriatally(into the caudate nucleus and putamen), or into the subthalamic nucleus(STN), for example optionally using the FDA-approved SMARTFLOW®neuroventricular cannula (SurgiVision, Inc.) specifically designed forclinical application, with or without the aid of the CLEARPOINT® systemto help the treating neurosurgeon(s) target and observe the delivery ofthe therapeutic agent in the brain (See, for example, San Sebastian, etal., 2014, Mol. Ther. Methods Clin. Dev. 3: 14049; See, for example,Feng and Maguire-Zeiss, 2010, CNS Drugs 24(3):177-192).

For example, during the surgery, two target points are determined in thesubstantia nigra pars compacta (SNpc) and ventral tegmental area (VTA)that are sufficiently separated from each other in dorsolateraldirections and identified on a magnetic resonance image. One burr holeis trepanned in each side of the cranial bone, through which the vectoris injected into the two target points via the two-track insertionroute. The AAV-vector-containing solution is prepared to a concentrationof 1.5×10¹² vector genome/ml, and 50 μl per point of the solution isinjected at 1 μl/min; each patient receives 3×10¹¹ vector genome of theAAV-vector construct.

Neutralizing antibody titers against AAV2 are determined by measuringβ-galactosidase activities in HEK293 cells transduced with 5×10³ vectorgenome/cell of AAV2 vectors expressing β-galactosidase in variousdilutions of sera.

PET

The AADC expression level in the substantia nigra are assessed on PETimaging with FMT six days before surgery and at one- and six-monthsafter gene transfer. All patients cease taking dopaminergic medications18 hours before PET and take 2.5 mg/kg of carbidopa orally one hourbefore FMT injection. Subsequently, 0.12 mCi/kg of FMT in saline isinfused into an antecubital vein, and a 90-minute dynamic acquisitionsequence is obtained. The PET and magnetic resonance imaging data areco-registered with a fusion processing program (Syntegra; Philips,Amsterdam, The Netherlands) to produce the fusion images.Radioactivities within volumes of interest drawn in the nigrostriatalpathway are calculated between 80 and 90 minutes after tracer injection.A change in nigrostriatal pathway FMT uptake from baseline to 24 weeksis assessed using the substantia nigra to striatal ratio ofradioactivity.

Statistical Analysis

Values at baseline and 6 months after gene transfer are compared usingStudent's t-test (paired analyses). A two-sided P value <0.05 is takento indicate significant differences. Two-way analysis of variance withBonferroni correction of P values is used for the short-durationresponse to levodopa. (See, for example, Muramatsu, et al., 2010, “Aphase I study of aromatic L-amino acid decarboxylase gene therapy forParkinson's disease.” Mol. Ther. 18:1731-1735).

Safety and tolerability of bilateral administration of AAV-vectorcompositions using real-time image-guided infusion into the brains ofParkinson's Disease subjects may be monitored for up to or after 9months post-surgery. Broad coverage of targeted areas (substantia nigrapars compacta (SNpc) and ventral tegmental area (VTA)) and widespreadAADC protein distribution in the striatum should be achieved withoutinducing any adverse effects.

Changes in Growth and Motor Skills:

The patients should gain weight and exhibit improvement in their motorscores after gene transfer, within a year, post-treatment. Weight willbe measured at 3 to 6 months after gene transfer. All patients initiallyshould have raw scores of zero on the Alberta Infant Motor Scale (AIMS)and very low raw scores for the Peabody Developmental Motor Scale,Second Edition (PDMS-II). After the gene transfer, all of the patientsshould show continuous increases in their raw scores on these twoscales, which indicates that their motor functions have improved. TheComprehensive Developmental Inventory for Infants and Toddlers (CDIIT)covers both cognition and motor development. All of the patients shouldshow low raw CDIIT scores before gene transfer, and the subsequentincrease in scores demonstrate improvement in both motor and cognitivefunctions.

Subjective Improvements after Gene Transfer

To document the symptoms that are more difficult to quantify, spouses,guardians or caretakers of the patients are asked to fill out aquestionnaire at the end of the study. The symptoms of the oculogyriccrises should lessen, and eye deviations and sleep disruptions, forexample, are some mild symptoms of the oculogyric crises that may remainafter gene therapy. Subjects may experience increased emotionalstability, and/or some improvements in sweating and hyperthermia (acommon manifestation of body temperature instability in hot weather).There should be no detectable abnormality in heart rate variability asassessed by 24-hour Holter monitoring either before or after genetransfer. Before gene therapy, patients that were bedridden and showedlittle spontaneous movement may exhibit less severe ptosis (drooping ofthe upper eyelid) one to two weeks after the gene transfer. According toprevious studies, dyskinesia may occur one month after gene transfer,but upon observation of a decrease in dyskinesia, motor developmentshould start (Hwu, W. L., et al., 2012. Gene therapy for aromaticL-amino acid decarboxylase deficiency. Sci. Transl. Med. Vol. 4,134ra61). Subjects may exhibit increased head control after threemonths, sitting with support after six to nine months, sitting up fromthe prone position after thirteen months, and holding toys and standingwith support sixteen months after the gene transfer, for example.Anti-AAV2 antibodies should be negative in the patients before genetherapy, and the titers may increase slightly after gene transfer.

PET Scans and CSF Analyses

PET scans and CSF analyses are completed for the treated patients. Sixmonths after gene transfer, PET scans should reveal that uptake of6-[18F] fluorodopa (FDOPA) increase from baseline in the combined (rightand left) treatment sites. The CSF analysis should reveal increases inthe levels of homovanillic acid (HVA, a metabolite of dopamine) and5-hydroxyindoleacetic acid (HIAA, a metabolite of serotonin). However,the levels of L-DOPA and 3-O-methyldopa may remain elevated (Hwu, W. L.,et al., 2012. Gene therapy for aromatic L-amino acid decarboxylasedeficiency. Sci. Transl. Med. Vol. 4, 134ra61).

Example 4. Administration of AADC Polynucleotides

AAV particle compositions are infused into the putamen of patientshaving Parkinson's Disease using the administration methods described inExample 3. The dose, number of patients and volume are outlined in Table4.

TABLE 4 Study Design Number Study No. of Patients Dose Volume 1 6  3 ×10¹¹ vg 100 ul per putamen 2 6  9 × 10¹¹ vg 300 ul per putamen 3 10 2.3× 10¹¹ vg 100 ul per putamen 4 10 7.5 × 10¹¹ vg 100 ul per putamen 5 57.5 × 10¹¹ vg 450 ul per putamen 6 Up to 20 1.4 × 10¹² vg Up to 900 ulper putamen 7 Up to 20 4.8 × 10¹² vg Up to 900 ul per putamen 8 Up to 208.8 × 10¹² vg Up to 900 ul per putamen

During the course of the study the safety and tolerability of theinfusion of the AADC polynucleotide-containing recombinantadeno-associated virus (AAV) vector compositions in human patientsdiagnosed with Parkinson's Disease is evaluated. Patients are evaluatedpreoperatively and monthly postoperatively for six months, usingmultiple measures, including the Global Systonia Scale (GDS) (seeComella, et al., 2003, Movement Disorders, 18(3):303-312), L-DOPAchallenge test, UPDRS scores, motor state diaries, and laboratory tests.Using diaries that separate the day into half-hour segments, thecaregivers of the patients will record their mobility during the fourdays before admission and for another four days at six months afteradmission to the study site. The patient caregivers are trained to ratesubject's condition as sleeping, immobile, mobile without troublesomedyskinesias, or mobile with troublesome dyskinesias. The total number ofhours spent in each of these categories is calculated, and thedifferences between the baseline and the six-month scores are comparedbetween the groups. The short-duration response to levodopa is evaluatedat baseline and 6 months after gene transfer; subjects take 100 mg oflevodopa orally with 25 mg benserazide after 20 hours withoutdopaminergic medication. Motor symptoms based on GDS and plasma levodopaconcentrations are assessed at baseline and 30 minutes, 1, 2, 3, and 4hours after levodopa intake (See, for example, Muramatsu, et al., 2010,“A phase I study of aromatic L-amino acid decarboxylase gene therapy forParkinson's disease.” z Mol. Ther. 18:1731-1735).

While the present disclosure has been described at some length and withsome particularity with respect to the several described embodiments, itis not intended that it should be limited to any such particulars orembodiments or any particular embodiment, but it is to be construed withreferences to the appended claims so as to provide the broadest possibleinterpretation of such claims in view of the prior art and, therefore,to effectively encompass the intended scope of the disclosure.

All publications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety. Incase of conflict, the present specification, including definitions, willcontrol. In addition, section headings, the materials, methods, andexamples are illustrative only and not intended to be limiting.

1. An aromatic L-amino acid decarboxylase (AADC) polynucleotidecomprising an AADC sequence region which has at least 95% identity toSEQ ID NO:
 979. 2. The AADC polynucleotide of claim 1, wherein the AADCsequence region comprises a promoter region, an enhancer region, amultiple cloning site (MCS) region and a polyadenylation (poly(A))signal region.
 3. The AADC polynucleotide of claim 2, wherein the AADCsequence region comprises at least one 5′ inverted terminal repeat (ITR)region and one 3′ ITR region.
 4. The AADC polynucleotide of claim 3,wherein one or more of the 5′ ITRs are located 5′ to the MCS region andone or more of the 3′ ITRs are located 3′ to the poly(A) signal.
 5. TheAADC polynucleotide of claim 3, wherein the AADC sequence regioncomprises a first exon region, a first intron region, a second intronregion and a second exon region.
 6. The AADC polynucleotide of claim 5,wherein the enhancer region and the promoter region are derived fromCMV.
 7. The AADC polynucleotide of claim 6, wherein the first exonregion is immediate-early 1 (ie1) exon 1 or fragments thereof, the firstintron region is ie1 intron 1 or fragments thereof, the second intronregion is human beta-globin (hBglobin) intron 2 or fragments thereof andthe second exon region is hBglobin exon 3 or fragments thereof.
 8. TheAADC polynucleotide of claim 2, wherein the poly(A) signal is derivedfrom human growth hormone.
 9. A plasmid or vector encoding the AADCpolynucleotide of any of claims 1-8.
 10. The plasmid or vector of claim9 which is or is derived from an adeno-associated virus (AAV).
 11. Arecombinant AAV virus comprising the AAV plasmid or vector of claim 10.12. The recombinant AAV virus of claim 11, comprising a capsid serotypeselected from the group consisting of AAV2, PHP.B, PHP.A, AAV1, AAV2G9,AAV3, AAV3a, AAV3b, AAV3-3, AAV4, AAV4-4, AAV5, AAV6, AAV6.1, AAV6.2,AAV6.1.2, AAV7, AAV7.2, AAV8, AAV9, AAV9.11, AAV9.13, AAV9.16, AAV9.24,AAV9.45, AAV9.47, AAV9.61, AAV9.68, AAV9.84, AAV9.9, AAV10, AAV11,AAV12, AAV16.3, AAV24.1, AAV27.3, AAV42.12, AAV42-1b, AAV42-2, AAV42-3a,AAV42-3b, AAV42-4, AAV42-5a, AAV42-5b, AAV42-6b, AAV42-8, AAV42-10,AAV42-11, AAV42-12, AAV42-13, AAV42-15, AAV42-aa, AAV43-1, AAV43-12,AAV43-20, AAV43-21, AAV43-23, AAV43-25, AAV43-5, AAV44.1, AAV44.2,AAV44.5, AAV223.1, AAV223.2, AAV223.4, AAV223.5, AAV223.6, AAV223.7,AAV1-7/rh.48, AAV1-8/rh.49, AAV2-15/rh.62, AAV2-3/rh.61, AAV2-4/rh.50,AAV2-5/rh.51, AAV3.1/hu.6, AAV3.1/hu.9, AAV3-9/rh.52, AAV3-11/rh.53,AAV4-8/r11.64, AAV4-9/rh.54, AAV4-19/rh.55, AAV5-3/rh.57, AAV5-22/rh.58,AAV7.3/hu.7, AAV16.8/hu.10, AAV16.12/hu.11, AAV29.3/bb.1, AAV29.5/bb.2,AAV106.1/hu.37, AAV114.3/hu.40, AAV127.2/hu.41, AAV127.5/hu.42,AAV128.3/hu.44, AAV130.4/hu.48, AAV145.1/hu.53, AAV145.5/hu.54,AAV145.6/hu.55, AAV161.10/hu.60, AAV161.6/hu.61, AAV33.12/hu.17,AAV33.4/hu.15, AAV33.8/hu.16, AAV52/hu.19, AAV52.1/hu.20, AAV58.2/hu.25,AAVA3.3, AAVA3.4, AAVA3.5, AAVA3.7, AAVC1, AAVC2, AAVC5, AAV-DJ,AAV-DJ8, AAVF3, AAVF5, AAVH2, AAVrh.72, AAVhu.8, AAVrh.68, AAVrh.70,AAVpi.1, AAVpi.3, AAVpi.2, AAVrh.60, AAVrh.44, AAVrh.65, AAVrh.55,AAVrh.47, AAVrh.69, AAVrh.45, AAVrh.59, AAVhu.12, AAVH6, AAVLK03,AAVH-1/hu.1, AAVH-5/hu.3, AAVLG-10/rh.40, AAVLG-4/rh.38, AAVLG-9/hu.39,AAVN721-8/rh.43, AAVCh.5, AAVCh.5R1, AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5,AAVCy.5R1, AAVCy.5R2, AAVCy.5R3, AAVCy.5R4, AAVcy.6, AAVhu.1, AAVhu.2,AAVhu.3, AAVhu.4, AAVhu.5, AAVhu.6, AAVhu.7, AAVhu.9, AAVhu.10,AAVhu.11, AAVhu.13, AAVhu.15, AAVhu.16, AAVhu.17, AAVhu.18, AAVhu.20,AAVhu.21, AAVhu.22, AAVhu.23.2, AAVhu.24, AAVhu.25, AAVhu.27, AAVhu.28,AAVhu.29, AAVhu.29R, AAVhu.31, AAVhu.32, AAVhu.34, AAVhu.35, AAVhu.37,AAVhu.39, AAVhu.40, AAVhu.41, AAVhu.42, AAVhu.43, AAVhu.44, AAVhu.44R1,AAVhu.44R2, AAVhu.44R3, AAVhu.45, AAVhu.46, AAVhu.47, AAVhu.48,AAVhu.48R1, AAVhu.48R2, AAVhu.48R3, AAVhu.49, AAVhu.51, AAVhu.52,AAVhu.54, AAVhu.55, AAVhu.56, AAVhu.57, AAVhu.58, AAVhu.60, AAVhu.61,AAVhu.63, AAVhu.64, AAVhu.66, AAVhu.67, AAVhu.14/9, AAVhu.t 19, AAVrh.2,AAVrh.2R, AAVrh.8, AAVrh.8R, AAVrh.10, AAVrh.12, AAVrh.13, AAVrh.13R,AAVrh.14, AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.20, AAVrh.21, AAVrh.22,AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.31, AAVrh.32, AAVrh.33, AAVrh.34,AAVrh.35, AAVrh.36, AAVrh.37, AAVrh.37R2, AAVrh.38, AAVrh.39, AAVrh.40,AAVrh.46, AAVrh.48, AAVrh.48.1, AAVrh.48.1.2, AAVrh.48.2, AAVrh.49,AAVrh.51, AAVrh.52, AAVrh.53, AAVrh.54, AAVrh.56, AAVrh.57, AAVrh.58,AAVrh.61, AAVrh.64, AAVrh.64R1, AAVrh.64R2, AAVrh.67, AAVrh.73,AAVrh.74, AAVrh8R, AAVrh8R A586R mutant, AAVrh8R R533A mutant, AAAV,BAAV, caprine AAV, bovine AAV, AAVhE1.1, AAVhEr1.5, AAVhER1.14,AAVhEr1.8, AAVhEr1.16, AAVhEr1.18, AAVhEr1.35, AAVhEr1.7, AAVhEr1.36,AAVhEr2.29, AAVhEr2.4, AAVhEr2.16, AAVhEr2.30, AAVhEr2.31, AAVhEr2.36,AAVhER1.23, AAVhEr3.1, AAV2.5T, AAV-PAEC, AAV-LK01, AAV-LK02, AAV-LK03,AAV-LK04, AAV-LK05, AAV-LK06, AAV-LK07, AAV-LK08, AAV-LK09, AAV-LK10,AAV-LK11, AAV-LK12, AAV-LK13, AAV-LK14, AAV-LK15, AAV-LK16, AAV-LK17,AAV-LK18, AAV-LK19, AAV-PAEC2, AAV-PAEC4, AAV-PAEC6, AAV-PAEC7,AAV-PAEC8, AAV-PAEC11, AAV-PAEC12, AAV-2-pre-miRNA-101, AAV-8h, AAV-8b,AAV-h, AAV-b, AAV SM 10-2, AAV Shuffle 100-1, AAV Shuffle 100-3, AAVShuffle 100-7, AAV Shuffle 10-2, AAV Shuffle 10-6, AAV Shuffle 10-8, AAVShuffle 100-2, AAV SM 10-1, AAV SM 10-8, AAV SM 100-3, AAV SM 100-10,BNP61 AAV, BNP62 AAV, BNP63 AAV, AAVrh.50, AAVrh.43, AAVrh.62, AAVrh.48,AAVhu.19, AAVhu.11, AAVhu.53, AAV4-8/rh.64, AAVLG-9/hu.39,AAV54.5/hu.23, AAV54.2/hu.22, AAV54.7/hu.24, AAV54.1/hu.21,AAV54.4R/hu.27, AAV46.2/hu.28, AAV46.6/hu.29, AAV128.1/hu.43, true typeAAV (ttAAV), UPENN AAV 10, Japanese AAV 10 serotypes, AAV CBr-7.1, AAVCBr-7.10, AAV CBr-7.2, AAV CBr-7.3, AAV CBr-7.4, AAV CBr-7.5, AAVCBr-7.7, AAV CBr-7.8, AAV CBr-B7.3, AAV CBr-B7.4, AAV CBr-E1, AAVCBr-E2, AAV CBr-E3, AAV CBr-E4, AAV CBr-E5, AAV CBr-e5, AAV CBr-E6, AAVCBr-E7, AAV CBr-E8, AAV CHt-1, AAV CHt-2, AAV CHt-3, AAV CHt-6.1, AAVCHt-6.10, AAV CHt-6.5, AAV CHt-6.6, AAV CHt-6.7, AAV CHt-6.8, AAVCHt-P1, AAV CHt-P2, AAV CHt-P5, AAV CHt-P6, AAV CHt-P8, AAV CHt-P9, AAVCKd-1, AAV CKd-10, AAV CKd-2, AAV CKd-3, AAV CKd-4, AAV CKd-6, AAVCKd-7, AAV CKd-8, AAV CKd-B1, AAV CKd-B2, AAV CKd-B3, AAV CKd-B4, AAVCKd-B5, AAV CKd-B6, AAV CKd-B7, AAV CKd-B8, AAV CKd-H1, AAV CKd-H2, AAVCKd-H3, AAV CKd-H4, AAV CKd-H5, AAV CKd-H6, AAV CKd-N3, AAV CKd-N4, AAVCKd-N9, AAV CLg-F1, AAV CLg-F2, AAV CLg-F3, AAV CLg-F4, AAV CLg-F5, AAVCLg-F6, AAV CLg-F7, AAV CLg-F8, AAV CLv-1, AAV CLv1-1, AAV Clv1-10, AAVCLv1-2, AAV CLv-12, AAV CLv1-3, AAV CLv-13, AAV CLv1-4, AAV Clv1-7, AAVClv1-8, AAV Clv1-9, AAV CLv-2, AAV CLv-3, AAV CLv-4, AAV CLv-6, AAVCLv-8, AAV CLv-D1, AAV CLv-D2, AAV CLv-D3, AAV CLv-D4, AAV CLv-D5, AAVCLv-D6, AAV CLv-D7, AAV CLv-D8, AAV CLv-E1, AAV CLv-K1, AAV CLv-K3, AAVCLv-K6, AAV CLv-L4, AAV CLv-L5, AAV CLv-L6, AAV CLv-M1, AAV CLv-M11, AAVCLv-M2, AAV CLv-M5, AAV CLv-M6, AAV CLv-M7, AAV CLv-M8, AAV CLv-M9, AAVCLv-R1, AAV CLv-R2, AAV CLv-R3, AAV CLv-R4, AAV CLv-R5, AAV CLv-R6, AAVCLv-R7, AAV CLv-R8, AAV CLv-R9, AAV CSp-1, AAV CSp-10, AAV CSp-11, AAVCSp-2, AAV CSp-3, AAV CSp-4, AAV CSp-6, AAV CSp-7, AAV CSp-8, AAVCSp-8.10, AAV CSp-8.2, AAV CSp-8.4, AAV CSp-8.5, AAV CSp-8.6, AAVCSp-8.7, AAV CSp-8.8, AAV CSp-8.9, AAV CSp-9, AAV.hu.48R3, AAV.VR-355,AAV3B, AAV4, AAV5, AAVF1/HSC1, AAVF11/HSC11, AAVF12/HSC12, AAVF13/HSC13,AAVF14/HSC14, AAVF15/HSC15, AAVF16/HSC16, AAVF17/HSC17, AAVF2/HSC2,AAVF3/HSC3, AAVF4/HSC4, AAVF5/HSC5, AAVF6/HSC6, AAVF7/HSC7, AAVF8/HSC8,AAVF9/HSC9, PHP.B (AAV-PHP.B), PHP.A (AAV.PHP.A), G2B-26, G2B-13,TH1.1-32, TH1.1-35, AAVPHP.B2, AAVPHP.B3, AAVPHP.N/PHP.B-DGT,AVPHP.B-EST, AAVPHP.BGGT, AAVPHP.B-ATP, AAVPHP.B-ATT-T, AAVPHP.B-DGT-T,AAVPHP.B-GGT-T, AAVPHP.B-SGS, AAVPHP.B-AQP, AAVPHP.B-QQP,AAVPHP.B-SNP(3), AVPHP.BSNP, AAVPHP.B-QGT, AAVPHP.B-NQT, AAVPHP.B-EGS,AAVPHP.B-SGN, AAVPHP.B-EGT, AAVPHP.B-DST, AAVPHP.B-DST, AAVPHP.B-STP,AAVPHP.BPQP, AAVPHP.B-SQP, AAVPHP.B-QLP, AAVPHP.B-TMP, AAVPHP.B-TTP,AAVPHP.S/G2A12, AAVG2A15/G2A3, AAVG2B4, and/or AAVG2B5, and variantsthereof.
 13. The recombinant AAV virus of claim 12, wherein the capsidserotype is AAV2.
 14. The recombinant AAV virus of claim 12, wherein thecapsid serotype is AAVrh10.
 15. The recombinant AAV virus of claim 12,wherein the capsid serotype is AAV9 (hu14).
 16. The recombinant AAVvirus of claim 12, wherein the capsid serotype is AAV-DJ.
 17. Therecombinant AAV virus of claim 12, wherein the capsid serotype isAAV9.47.
 18. The recombinant AAV virus of claim 12, wherein the capsidserotype is AAV-DJ8.
 19. The AADC polynucleotide of claim 1, wherein theAADC sequence region has at least 99% identity SEQ ID NO:
 979. 20. Apharmaceutical composition comprising an adeno-associated virus (AAV)particle, said AAV particle comprising an AAV capsid and a vectorgenome, said vector genome comprising at least one AADC sequence regionwith at least 95% identity to SEQ ID NO:
 900. 21. The pharmaceuticalcomposition of claim 20, wherein the AADC sequence region has at least99% identity SEQ ID NO:
 979. 22. The pharmaceutical composition of claim20, wherein at least 70% of the AAV particles contain a vector genome.23. A method of reducing periods of dyskinesia in a subject comprisingadministering to said subject the pharmaceutical composition of claims20-22.
 24. The method of claim 23, wherein the decrease in periods ofdyskinesia is shown by improvement in off time and motor fluctuations byat least 30%.
 25. The method of claim 24, wherein the improvement lastsfor at least 6 hours.
 26. The method of any of claims 23-26, wherein thesubject has Parkinson's Disease.
 27. A method of improving thesleep-wake cycle of a subject comprising administering to said subjectthe pharmaceutical composition of claims 20-22.
 28. The method of claim27, where the subject's amount of rapid eye movement (REM) sleep isdecreased as compared to the subject's amount of REM sleep prior toadministration of the pharmaceutical composition.
 29. The method ofclaim 27, wherein the subject's amount of non-REM (NREM) sleep isincreased as compared to the subject's amount of REM sleep prior toadministration of the pharmaceutical composition.
 30. The method of anyof claims 27-29, wherein the subject has Parkinson's Disease.
 31. Amethod of treating a sleep disorder of a subject comprisingadministering to said subject the pharmaceutical composition of claims20-22.
 32. The method of claim 31, wherein the sleep disorder isinsomnia.
 33. The method of any of claim 31 or 32, wherein the subjecthas Parkinson's Disease.
 34. A method of increasing the level of AADCprotein in a subject comprising administering the pharmaceuticalcomposition of claims 20-22.